ALBERT

Description: 〈p〉Publication date: Available online 28 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Jiwon Woo, Haewon Kang, Eunice Yoojin Lee, Sangkyu Park, Yong Eun Cho〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Recent developments in tissue clearing methods such as CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging/Immunostaining/In situ hybridization-compatible Tissue hYdrogel) have allowed for the three-dimensional analysis of biological structures in whole, intact tissue, providing greater understanding of spatial relationships and biological circuits. Nonetheless, studies have reported issues with maintaining structural integrity and preventing tissue disintegration, preventing the wide application of these techniques to fragile tissues such as developing embryos. Here, we present optimized passive clearing techniques, mPACT-A, that improve tissue rigidity without the expense of optical transparency. We also present a further modified mPACT-A protocol that is specifically optimized for handling mouse embryos, which are small and fragile, such that they easily dismantle when processed via established tissue clearing methods. We demonstrate proof-of-concept by investigating the expression of two relatively understudied PRDM proteins, PRDM7 and PRDM12, in intact cleared mouse embryos at various stages of development. We observed strong PRDM7 and PRDM12 expression in the developing mouse nervous system, suggestive of potential roles in neural development that will be tested in future functional studies.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 28 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Tamaki Izume, Hirotake Miyauchi, Wataru Shihoya, Osamu Nureki〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Sarafotoxins (SRTXs) are endothelin-like peptides extracted from snake venom. SRTXs stimulate the endothelin ET〈sub〉A〈/sub〉 and ET〈sub〉B〈/sub〉 receptors and enhance vasoconstriction, followed by left ventricular dysfunction and bronchoconstriction. SRTXs include four major isopeptides, S6a-d, with different subtype selectivities. Here, we report the crystal structure of the human ET〈sub〉B〈/sub〉 receptor in complex with the non-selective sarafotoxin S6b at 3.0 Å resolution. This structure reveals the similarities and differences between the binding modes of the endothelins and S6b. Moreover, molecular dynamics simulations based on the S6b-bound receptor provides structural insight into the subtype selectivity of the sarafotoxins. Our study clarifies the recognition mechanism of the endothelin-like peptide families.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 28 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Jing Wu, Yuxing Zhao, Zhen Fan, Qiunan Chen, Jinliang Chen, Yue Sun, Xushun Jiang, Qian Xiao〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Diabetes mellitus is a metabolic disorder that can lead to blood-brain barrier (BBB) disruption and cognitive decline. However, the mechanisms of BBB breakdown in diabetes are still unclear. Soluble epoxide hydrolase (sEH) is an enzyme that degrades epoxyeicosatrienoic acids (EETs), which have multiple protective effects on vascular structure and functions. In the current study, we showed increased vascular permeability of the BBB, which was accompanied by upregulation of sEH and downregulation of 14,15-EET. Moreover, the sEH inhibitor t-AUCB restored diabetic BBB integrity 〈em〉in vivo〈/em〉, and 14,15-EET prevented ROS accumulation and MEC injury 〈em〉in vitro〈/em〉. t-AUCB or 14,15-EET treatment provoked AMPK/HO-1 activation under diabetic conditions 〈em〉in vivo〈/em〉 and 〈em〉in vitro.〈/em〉 Thus, we suggest that decreased EET degradation by sEH inhibition might be a potential therapeutic approach to attenuate the progression of BBB injury in diabetic mice via AMPK/HO-1 pathway activation.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 27 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Nikolai Jaschke, Alexander Kleymann, Lorenz C. Hofbauer, Andy Göbel, Tilman D. Rachner〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉 〈p〉Advanced stages of breast cancer are frequently complicated by bone metastases which cause substantial cancer-related morbidity and mortality. The Wnt-signaling antagonist Dickkopf-1 (DKK-1) has emerged as a crucial factor in the development and progression of osteolytic bone metastases. Although several signaling pathways have been implicated in promoting DKK-1 production in breast cancer cells, pharmacological interventions that interfere with tumor DKK-1 synthesis still remain scarce.〈/p〉 〈p〉In the current study, using an unbiased approach, we identified the small molecule Dorsomorphin as a potent suppressor of DKK-1 in several breast cancer cell lines (MDA-MB-231, MDA-Bone, MDA-MET and MCF7, respectively). Here, Dorsomorphin suppressed DKK-1 mRNA and protein production by 70 and 90%, respectively (p 〈0.001). Whereas bone morphogenic protein (BMP)- and AMP activated protein kinase (AMPK)-signaling are two well-established targets of Dorsomorphin, we show that neither pathway is essentially involved in facilitating its inhibitory effects on DKK-1.〈/p〉 〈p〉In summary, we identified Dorsomorphin as a potent pharmacological inhibitor of DKK-1 production in breast cancer cells. Whether Dorsomorphin reflects a valuable therapeutic agent in breast cancer warrants further investigations.〈/p〉 〈/div〉 〈/div〉

Description: 〈p〉Publication date: 19 February 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications, Volume 522, Issue 4〈/p〉 〈p〉Author(s): 〈/p〉

Description: 〈p〉Publication date: Available online 25 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Kun Dong, Qiang Du, Xiao Cui, Peiqi Wan, Christof Kaltenmeier, Jing Luo, Bing Yan, Yihe Yan, David A. Geller〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Hepatocellular carcinoma (HCC) tumors evade death in part by downregulating expression of the tumor suppressor gene Interferon regulatory factor-1 (IRF-1). However, the molecular mechanisms accounting for IRF-1 suppression in HCC have not been well described. In this study, we identified a novel microRNA-301a (miR-301a) binding site in the 3′-untranslated region (3′- UTR) of the human IRF-1 gene and hypothesized a functional role for miR-301a in regulating HCC growth. We show that miR-301a is markedly upregulated in primary HCC tumors and HCC cell lines, while IRF-1 is down-regulated in a post-transcriptional manner. MiR-301a regulates basal and inducible IRF-1 expression in HCC cells with an inverse relationship between miR-301a and IRF-1 expression in HCC cells. Chronic hypoxia induces miR-301a in HCC in vitro and decreases IRF-1 expression. Finally, miR-301a inhibition increases apoptosis and decreases HCC cell proliferation. These findings suggest that targeting of IRF-1 by miR-301a contributes to the molecular basis for IRF-1 downregulation in HCC and provides new insight into the regulation of HCC by miRNAs.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 24 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Takuto Shimizu, Yoshimichi Kozuka, Mayumi Kusano, Masaki Nagane, Tadashi Yamashita, Naomi Hachiya〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉In many neurodegenerative diseases, mitochondria are actively involved in the onset and/or progression of diseases because the energy depletion of the neuronal cells directly leads to the dysfunction and degeneration of cells. In the case of prion diseases, mitochondrial involvement has been reported recently and evidence that prion protein (PrP) is localized in mitochondria is increasing. Despite these findings, the precise molecular mechanism by which PrP targets mitochondria remains unclear. PrP is a secretory protein and does not have a pre-sequence that targets the mitochondria, therefore, we thought that there was a covert signal in the amino acid sequence of PrP. To find the sequence, we constructed various GFP-fused PrP-truncations and colocalization with mitochondria was verified by live-cell imaging. Consequently, we found that 18 amino acids, PrP (122–139), are indispensable for the mitochondrial targeting of PrP. In addition, fluorescent microscopy observation revealed that PrP-localized mitochondria were accumulated at the perinuclear region in neuronal cells such as mouse neuroblastoma Neuro2a (N2a) and prion persistent infection N2a strain (ScN2a), anterograde movement of the mitochondria toward the cell membrane was completely inhibited because of the stacking of PrP on the outer membrane. The cristae formation of perinuclear accumulated mitochondria was disappeared indicating the reduced mitochondrial activity. Surprisingly, PrP-dependent mitochondrial perinuclear accumulation was specifically occurred on neuronal cells, whereas in epithelial HeLa cells and fibroblast COS-7 cells, no perinuclear accumulation observed even after the mitochondrial targeting of PrP.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 25 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Natalia Putkaradze, Lisa König, Lars Kattner, Michael C. Hutter, Rita Bernhardt〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Vitamin D2 is a form of vitamin D derived from mushrooms and plants which is structurally modified in the body due to the action of several enzymes. The resulting metabolites represent important compounds with potential bioactive properties. However, they are poorly studied and their availability is mostly limited. In order to identify new enzymes capable of producing vitamin D2 metabolites, we investigated a bacterial P450 monooxygenase, CYP109E1, which was previously shown to be a vitamin D3 hydroxylase. It was found that CYP109E1 catalyzes a vitamin D2 two-step hydroxylation at positions C24 and C25 resulting in the generation of 24〈em〉(R)〈/em〉,25-diOH VD2. Interestingly, the enzyme showed high selectivity towards vitamin D2, whereas it showed an unselective product pattern for the structurally similar vitamin D3. Our docking results for vitamin D2 and D3 revealed favorable hydroxylation positions for both substrates and suggested an explanation for the high selectivity of CYP109E1 towards vitamin D2. In addition, we established a whole-cell biocatalyst expressing CYP109E1 in 〈em〉Bacillus megaterium〈/em〉 to produce 24〈em〉(R)〈/em〉,25-diOH VD2 and a production yield of 12.3 ± 1.2 mg/L was obtained after 48 h. To the best of our knowledge, this is the first report on the generation of 24〈em〉(R)〈/em〉,25-diOH VD2 by a microbial biocatalyst allowing a low-cost and eco-friendly production of this pharmaceutically interesting and expensive metabolite from the relatively cheap substrate, VD2.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 25 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Baosheng Wang, Jingjing Men, Chunmei Wang, Xiaoming Hou, Feng Zhao〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Successful induction of milk protein synthesis relies on prolactin/STAT5. In mice, both laminin and β1 integrin were necessary for STAT5 activity induced by prolactin treatment, resulting in transcriptional activation of β-casein. However, the mechanism by which β1 integrin increases the bovine milk protein synthesis is not well known. In order to display the crosstalk between integrin signaling and lactogenic signaling, we investigated the mechanism by which laminin mediated lactogenic effects via interaction with β1 integrin on bovine mammary epithelial cells (BMECs). Therefore, localization of β1 integrin was examined by immunofluorescence. The mRNA and protein expression levels were determined by quantitative real-time PCR and western blotting. The results showed that β1 integrin were detected in basal mammary cells and basal membrane surface of adherent BMECs. However, basal distribution of β1 integrin was not sufficient to increase β-casein synthesis in the absence of integrin activation by laminin. A lactogenic hormone cocktail of insulin, hydrocortisone, and prolactin stimulated overall lactogenic effects, including upregulated expression of β1 integrin, activation of prolactin/STAT5 signaling, and consequent increase of β-casein synthesis. In response to a 24 h prolactin treatment, the abundance of STAT5, β1 integrin, and β-casein in BMECs with laminin was higher compared to that with a control substratum. Meanwhile, laminin-dependent lactogenic effects were inhibited by blocking β1 integrin function, resulting in attenuated STAT5 activity and decreased β-casein synthesis. These results indicated that β1 integrin was a key mediator of the laminin-dependent prolactin/STAT5 signaling, which regulated the sustained STAT5 activity necessary for β-casein expression in BMECs.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 25 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Zhengdong Chen, Rengong Zhuo, Yun Zhao, Lichao Yang, Yu Zhou, Xiaoling Cheng, Lu Peng, Xin Jin, Yiqing Wang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉 〈h6〉Background〈/h6〉 〈p〉Atherosclerotic plaque rupture is the major trigger of acute cardiovascular risk events, and manipulation of M1/M2 macrophage homeostasis is an effective strategy for regulating atherosclerotic plaque stability. This study was aimed to illuminate the effects of oleoylethanolamide (OEA) on macrophage polarization and plaque stability.〈/p〉 〈/div〉 〈div〉 〈h6〉Methods〈/h6〉 〈p〉Macrophages derived from THP-1 were treated with OEA followed by LPS/IFN-γ, and the markers of M1, M2 macrophages were monitored by western blot, real-time PCR and immunofluorescence staining. The effect of OEA on macrophage polarization in the arch of aortic arteries was tested by immunofluorescence staining and western blot, and the plaque stability was completed by Masson’s trichrome and hematoxylin and eosin (HE) in apolipoprotein E (ApoE)〈sup〉−/−〈/sup〉 mice.〈/p〉 〈/div〉 〈div〉 〈h6〉Results〈/h6〉 〈p〉OEA treatment enhanced the expression of two classic M2 macrophage markers, macrophage mannose receptor (CD206) and transforming growth factor (TGF-β), while the expression of iNOS (M1 macrophages) was decreased in THP-1-derived macrophages. Blocking of PPARα using siRNA and inhibition of AMP-activated protein kinase (AMPK) by its inhibitor compound C attenuated the OEA-induced expression of M2 macrophage markers. In addition, OEA significantly suppressed M1, promoted M2 macrophage polarization, increased collagen content and decreased necrotic core size in atherosclerotic plaques in ApoE〈sup〉−/−〈/sup〉 mice, which were linked with the expression of PPARα.〈/p〉 〈/div〉 〈div〉 〈h6〉Conclusions〈/h6〉 〈p〉OEA improved atherosclerotic plaque stability through regulating macrophage polarization via AMPK-PPARα pathway.〈/p〉 〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 26 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Yasuyuki Momoi, Akihiko Nishikimi, Guangwei Du, Tohru Kataoka, Koko Katagiri〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Integrin activation by Rap1-GTP is pivotal for lymphocyte trafficking. In this study, we show the phosphatidic acid (PA)-dependent membrane distribution of RA-GEF-1 and -2 (also known as Rapgef2 and 6), which are guanine nucleotide exchange factors for Rap1, plays important roles in lymphocyte migration. RA-GEF-1 associates with PA through 919–967 aa within CDC25 homology domain, and the deletion of this region of RA-GEF-1 inhibits chemokine-dependent migration. Chemokine stimulation induces temporal production of PA on the plasma membrane, which is not necessary for Rap1 activation, but the translocation of RA-GEFs. Thus, chemokine-dependent generation of PA is critical for lymphocyte migration through membrane localization of RA-GEFs.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 26 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Hirofumi Nohara, Ryunosuke Nakashima, Shunsuke Kamei, Haruka Fujikawa, Keiko Ueno-Shuto, Taisei Kawakami, Yuka Eto, Mary Ann Suico, Jian-Dong Li, Hirofumi Kai, Tsuyoshi Shuto〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Glucagon-like peptide-1 (GLP-1) is a gastrointestinal hormone that stimulates glucose-mediated insulin production by pancreatic beta cells. It is also associated with protective effects in multiple tissues. GLP-1 receptor is highly expressed in pulmonary tissue, hinting possible pulmonary delivery of GLP-1 drugs. However, little is known about the role of GLP-1 signaling in the lung, especially in mucus hypersecretory obstructive lung diseases. Here, we showed that treatment with exendin-4, a clinically available GLP-1 receptor agonist, up-regulates mucin expression in normal airway epithelial cells and in the lung of normal mice, indicating mucus stimulatory effect of GLP-1 under physiological condition. Exendin-4 also increased mucin expression in 〈em〉in vitro〈/em〉 cellular and 〈em〉in vivo〈/em〉 murine models of obstructive lung diseases 〈em〉via〈/em〉 the activation of p38 MAP kinase. Notably, mucin induction 〈em〉in vivo〈/em〉 exacerbated key pulmonary abnormalities including emphysematous phenotypes, implying that GLP-1 signaling in the lung is detrimental under pulmonary obstructive condition. Another GLP-1 receptor agonist liraglutide had similar induction of mucin. Together, our studies not only demonstrate novel physiological and pathological roles of GLP-1 in the lung but may also caution against the clinical use of inhaled GLP-1 receptor agonists in the patients with obstructive lung diseases.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X20301509-fx1.jpg" width="285" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: Available online 26 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Yuya Nishimura, Yota Fukuda, Toya Okonogi, Soichiro Yoshikawa, Hajime Karasuyama, Naomi Osakabe, Yuji Ikegaya, Takuya Sasaki, Takahiro Adachi〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The brain–gut axis which is an interaction between recognition and emotion and the gut sensory system for food and microbiota is important for health. However, there is no real-time monitoring system of the brain and the gut simultaneously so far. We attempted to establish a dual real-time monitoring system for the brain–gut axis by a combination of intravital Ca〈sup〉2+〈/sup〉 imaging of the gut and electroencephalogram. Using a conditional Yellow Cameleon 3.60 expression mouse line, we performed intravital imaging of the gut, electrophysiological recordings of the vagus nerve, and electroencephalogram recordings of the various cortical regions simultaneously upon capsaicin stimuli as a positive control. Upon capsaicin administration into the small intestinal lumen, a simultaneous response of Ca〈sup〉2+〈/sup〉 signal in the enteric nervous system and cortical local field potentials (LFPs) was successfully observed. Both of them responded immediately upon capsaicin stimuli. Capsaicin triggered a significant increase in the frequency of vagus nerve spikes and a significant decrease in the slow-wave power of cortical LFPs. Furthermore, capsaicin induced delayed and sustained Ca〈sup〉2+〈/sup〉 signal in intestinal epithelial cells and then suppressed intestinal motility. The dual real-time monitoring system of the brain and the gut enables to dissect the interaction between the brain and the gut over time with precision.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 23 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Xi Wang, Kai Li, Jialing Tang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Recent studies highlighted the significance of chronic inflammation, which is mediated in part by toll-like receptors 4 (TLR4), in islet β cell dysfunction by high-glucose exposure. However, about it is unclear whether islet β cell dysfunction in response to high glucose is associated with TLR4. This investigation was designed to address the effect of TLR4 deficiency on insulin secretion in mice in response to acute intravenous glucose load. Hyperglycemic clamp was used to impair insulin secretion, and intraperitoneal glucose tolerance test was carried out to analyze insulin secretion function of islet β cells. Our results showed that TLR4 deficiency repressed insulin secretion impairment in response to acute intravenous glucose load. Compared to wild-type mice, TLR4〈sup〉−/−〈/sup〉 mice did not exhibit increase of IL-1β and TNF-α level in plasma and pancreatic tissue in response to acute intravenous load of high glucose. However, recombinant IL-1β or TNF-α administration restored insulin secretion impairment induced by high glucose in TLR4〈sup〉−/−〈/sup〉 mice. Taken together, our results demonstrated that TLR4 activation and subsequent IL-1β and TNF-α production contribute to islet β cell dysfunction in mice in response to acute intravenous load of high glucose, which may provide a theoretical basis for diabetes complication improvement by physical exercise.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 26 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Jiajia Wang, Jia Guo, Bo Che, Mingxing Ouyang, Linhong Deng〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Extracellular Matrix (ECM) assembly and remodeling are critical physiological events 〈em〉in vivo〈/em〉, and abnormal ECM assembly or remodeling is related to pathological conditions such as osteoarthritis, fibrosis, cancers, and genetic diseases. ECM assembly/remodeling driven by cells represents more physiological processes. Collagen I (COL) is very abundant in tissues, which assembly/remodeling is mediated by biochemical and mechanical factors. How cells regulate COL assembly biomechanically still remains to be well understood. Here we used fluorescent COL in the medium to study how cells assembled ECM which represents more physiological structures. The results showed that MDCK cells actively recruited COL from the medium and helped assemble the fibers, which in turn facilitated cell branching morphogenesis, both displaying highly spatial associations and mutual dependency. Inhibition of cellular contraction force by ROCK and Myosin II inhibitors attenuated but did not block the COL fiber formation, while cell motion showed high consistency with the fiber assembly. Under ROCK or Myosin II inhibition, further analysis indicated high correlation between local cell movement and COL fiber strength as quantified from different regions of the same groups. Blocking cell motion by actin cytoskeleton disruption completely inhibited the fiber formation. These suggest that cell motion coordinated COL fiber assembly from the medium, possibly through generated strain on deposited COL to facilitate the fiber growth.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 23 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Kyung Min Kim, Ho Taek Oh, Gi Don Yoo, Jun-Ha Hwang, Areum Oh, Eun Sook Hwang, Jeong-Ho Hong〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Ultraviolet (UV) irradiation induces the proliferation and differentiation of keratinocytes in the basal layer of the epidermis, which increases epidermal thickness in skin regeneration. However, the mechanism underlying this phenomenon is not yet known in detail. In this study, we aimed to demonstrate that the transcriptional coactivator with PDZ-binding motif (TAZ) stimulates epidermal regeneration by increasing keratinocyte proliferation. During epidermal regeneration, TAZ is localized in the nucleus of keratinocytes of the basal layer and stimulates epidermal growth factor receptor (EGFR) signaling. TAZ depletion in keratinocytes decreased EGFR signaling activation, which delays epidermal regeneration. Interestingly, TAZ stimulated the transcription of amphiregulin (AREG), a ligand of EGFR, through TEAD-mediated transcriptional activation. Together, these results show that TAZ stimulates EGFR signaling through AREG induction, suggesting that it plays an important role in epidermal regeneration.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 23 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Tyler L. Perfitt, Philip E. Stauffer, Keeley L. Spiess, Roger J. Colbran〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Protein-protein interactions can be modulated by phosphorylation of either binding partner, thereby altering subcellular localization and/or physiological function. Shank3, a master postsynaptic scaffolding protein that controls the developmental maturation of excitatory synapses, was recently shown to be phosphorylated by Protein Kinase A (PKA) at Ser685 〈em〉in vivo〈/em〉. Mutation of Shank3 Ser685 was shown to modulate the binding of Abelson interactor 1 (ABI1), a component of the WAVE regulatory complex for actin remodeling, but a direct effect of Ser685 phosphorylation on ABI1 binding was not investigated. Here, we demonstrate that Ca〈sup〉2+〈/sup〉/calmodulin-dependent protein kinase II alpha (CaMKIIα) also phosphorylates Shank3 at Ser685. Mutation of Ser685 to phospho-null alanine (S685A) prevented both CaMKIIα and PKA phosphorylation of a GST-Shank3 fusion protein. The co-immunoprecipitation of ABI1 with Shank3 from HEK293 cell extracts is reduced by mutation of Ser685 to either Ala or Asp. However, pre-phosphorylation of GST-Shank3 by purified CaMKIIα significantly increased binding of ABI1, and this effect was abrogated by Ser685 to Ala mutation in GST-Shank3. Taken together, our data suggest that neuronal ABI1-Shank3 interactions may be convergently regulated by Shank3 Ser685 phosphorylation in response to both Ca〈sup〉2+〈/sup〉 and cAMP signaling, potentially modulating dendritic spine morphology.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 23 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Chang Su, Tinghong Ming, Yan Wu, Qinqin Jiang, Hengshang Huan, Chenyang Lu, Jun Zhou, Ye Li, Hongying Song, Xiurong Su〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Ferritins are ubiquitous iron-binding proteins that are mainly related to iron storage, detoxification and innate immunity. Here, we present the crystal structure of a marine invertebrate ferritin from 〈em〉Sinonovacula constricta〈/em〉 at a resolution of 1.98 Å. The 〈em〉S. constricta〈/em〉 ferritin (ScFer) possessed some structural similarities with vertebrate ferritins, and they shared a well-conserved architecture composed of five α-helical bundles that assembled into a cage-like structure with 24-subunits. The structure of ScFer also showed iron binding sites in the 3-fold channel, ferroxidase center, and putative nucleation sites. Further, electrostatic potential calculations suggested that the electrostatic gradient of the 3-fold channel could provide a guidance mechanism for iron entering the ferritin cavity.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 23 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Arif Md. Rashedul Kabir, Kazuki Sada, Akira Kakugo〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Microtubule is the most rigid component of eukaryotic cytoskeleton that plays pivotal roles in many important cellular events. Microtubules are known to undergo bending or buckling in cells which often results in breaking of this cytoskeletal protein filament. Various cellular events such as cell migration, chromosome segregation, etc. are dependent on the buckling induced breaking of microtubules. However, the reason behind the breaking of buckled microtubules in cell has remained obscure yet. In this work, we have demonstrated breaking of microtubules on a 2D elastic medium by applying compressive stress. The applied compressive stress caused buckling of the microtubules which ultimately resulted in their breaking. We show that breaking of the buckled microtubules cannot be accounted for by considering the changes in curvature of the microtubules due to mechanical deformation. Our results confirm that, it is the interaction of kinesin, a microtubule-associated motor protein, with microtubules which plays the key role in breaking of the buckled microtubules on the 2D elastic medium. The breaking of buckled microtubules is ascribed to decrease in rigidity of microtubules upon interaction with kinesins. This work for the first time confirms the involvement of a microtubule-associated motor protein in breaking of microtubules under compressive stress, which will help further clarify the mechanism of breaking of buckled microtubules in cells and its significance in the cellular events.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉Biomolecular motor kinesin mediates breaking microtubules (MT) under compressive stress by decreasing the rigidity of microtubules.〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X20301510-fx1.jpg" width="145" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: Available online 23 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Sunmi Kang, Hyuk Nam Kwon, Soeun Kang, Sunghyouk Park〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Isocitrate dehydrogenase (IDH) mutations are found in low-grade gliomas, and the product of the IDH mutant (MT), 2-hydroxyglutarate (2-HG), is the first known oncometabolite. However, the roles of the IDH wild type (WT) in high-grade glioblastoma, which rarely has the IDH mutation, are still unknown. To investigate possible pathways related to IDH WT in gliomas, we carried out bioinformatics analysis, and found that IDH1 has several putative calmodulin (CaM) binding sites. Pull-down and quantitative dissociation constant (Kd) measurements using recombinant proteins showed that IDH1 WT indeed binds to CaM with a higher affinity than IDH1 R132H MT. This biochemical interaction was demonstrated also in the cellular environment by immunoprecipitation with glioblastoma cell extracts. A synthetic peptide for the suggested binding region interfered with the interaction between CaM and IDH1, confirming the specificity of the binding. Direct binding between the synthetic peptide and CaM was observed in an NMR binding experiment, which additionally revealed that the peptide initially binds to the C-lobe of CaM. The physiological meaning of the CaM-IDH1 WT binding was shown with trifluoperazine (TFP), a CaM antagonist, which disrupted the binding and inhibited survival and migration of glioblastoma cells with IDH1 WT. As CaM signaling is activated in glioblastoma, our results suggest that IDH1 WT may be involved in the CaM-signaling pathway in the tumorigenesis of high-grade gliomas.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 23 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Alexander McPherson〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉X-ray intensities extending to 1.4 Å resolution were collected on the P6〈sub〉3〈/sub〉 hexagonal crystal form of canavalin, and extended to 1.9 Å for the orthorhombic C222〈sub〉1〈/sub〉 crystals. Structure determination of a new crystal form of canavalin having space group P2〈sub〉1〈/sub〉2〈sub〉1〈/sub〉2〈sub〉1〈/sub〉 is reported as well. Both the N and C terminal cupin domains contained identifiable ligands. For hexagonal crystals, in the cavity of the C terminal cupin, a molecule of benzoic acid was found, bound through carboxyl oxygens to Histidine 297, asparagine 284 and Arginine 376. The benzene ring was immersed in a cluster of at least 8 hydrophobic amino acid side chains. The N terminal cupin contained a molecule of citrate. Benzoic acid was also found to be present in the C terminal cupins of in the C222〈sub〉1〈/sub〉 and P2〈sub〉1〈/sub〉2〈sub〉1〈/sub〉2〈sub〉1〈/sub〉 crystal forms. In rhombohedral crystals, the C terminal cupin domain appeared to be occupied by a phosphate ion, but this was ambiguous. In cubic crystals, both domains were vacant. The N terminal cupin domains of canavalin in the P2〈sub〉1〈/sub〉2〈sub〉1〈/sub〉2〈sub〉1〈/sub〉 and rhombohedral crystals were also vacant, but the N terminal cupin domain of the C222〈sub〉1〈/sub〉 crystals contained a ligand whose identity is uncertain, but which has been modeled as HEPES buffer. A possible physiological role for the ligands and their complexes with canavalin is considered.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉Benzoic acid, an important plant metabolite, is shown as it is bound in a cupin domain of a vicilin protein, canavalin, from Canavalia ensiformis. The binding suggests that vicilin proteins may serve additional biochemical roles other than as storage proteins.〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X20301765-fx1.jpg" width="312" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: Available online 23 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Tomotaka Kurihara, Toru Asahi, Naoya Sawamura〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Cereblon (〈em〉CRBN〈/em〉) was identified as a gene that causes intellectual disabilities. The encoded CRBN protein, containing 442 amino acids, is located in several organs. Cytosolic CRBN was reported to mainly act as a component of the E3 ubiquitin ligase complex. CRBN is one of the substrate receptors of the E3 ubiquitin ligase complex and promotes the degradation of targeted proteins. Studies have reported that CRBN recognizes the C-terminal region of the amyloid precursor protein (APP), a protein known for its involvement in the development of Alzheimer’s disease. Although CRBN may interact with the C-terminal region of APP in mice, the CRBN-mediated degradation mechanism of human APP remains unclear. Here, we analyzed the CRBN-mediated degradation mechanism of human APP via the ubiquitin-proteasome system. Immunoprecipitation experiments showed that CRBN interacts with human full-length APP via its C-terminal region. Next, we examined CRBN-mediated degradation of APP in the ubiquitin-proteasome system. CRBN recognizes Lys〈sup〉751〈/sup〉 in human APP and ubiquitinates it in SH-SY5Y cells. Overexpression of CRBN decreased wild-type APP expression levels. In contrast, the expression level of K751R APP remained unchanged by CRBN overexpression, while knockdown of endogenous CRBN increased APP levels. As such, our results suggest that CRBN ubiquitinates Lys〈sup〉751〈/sup〉 of human APP thereby degrading it via the ubiquitin-proteasome system.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 23 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Dong Hee Kim, Eun Ji Kim, Do Hee Kim, Seung Woo Park〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Dishevelled-associated antagonist of beta-catenin 2 (〈em〉Dact2〈/em〉) is involved in the regulation of intracellular signaling pathways during development. It negatively regulates the Nodal signaling pathway, possibly by promoting lysosomal degradation of Nodal receptors such as TGFBR1, and plays an inhibitory role during the re-epithelialization of skin wounds by attenuating transforming growth factor-β signaling. 〈em〉Dact〈/em〉2 is known to act as a functional tumor suppressor in colon cancer; reduced 〈em〉Dact2〈/em〉 can promote liver cancer progression and suppress gastric cancer proliferation, invasion, and metastasis by inhibiting Wnt signaling. Zebrafish is used as a model of cancer biology because it shows similar tumorigenesis and morphogenesis as in humans and gene manipulation in this organism is possible. This study was performed to explore phenotypic changes in 〈em〉Dact2〈/em〉 knockout zebrafish and investigate the function of 〈em〉Dact2〈/em〉. A 10-base pair deletion 〈em〉Dact2〈/em〉 knockout zebrafish was prepared using the CRISPR-Cas9 genome editing system. 〈em〉Dact2〈/em〉 knockout enhanced the expression of the 〈em〉MMP2〈/em〉 and 〈em〉MMP9〈/em〉 genes, which are related to tumor invasion and migration, and the 〈em〉Snail〈/em〉, 〈em〉VEGF〈/em〉, and 〈em〉ZEB〈/em〉 genes, which are related to epithelial-mesenchymal transition (EMT). The absence of 〈em〉Dact2〈/em〉 also resulted in hyperplasia of the gastrointestinal epithelium, fibrosis in the pancreas and liver, increased proliferation of the pancreatic and hepatic bile ducts, and invasive proliferation into the pancreas. A wound healing assay confirmed that the absence of 〈em〉Dact2〈/em〉 enhanced EMT, thus accelerating wound healing. This study suggests that a loss of function of 〈em〉Dact2〈/em〉 impacts EMT-related gene regulation and tumor generation in a zebrafish knockout model, which is a useful model for exploring the mechanisms of these processes.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 23 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Shaohai Wang, Yuan Li, Si Sun, Jing Cai, Jin Cao〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Decreased miR-335 has been reported in a variety of cancers. We previously showed that miR-335 played an important role in ovarian cancer metastasis and prognosis. However, miR-335 is down-regulated in ovarian cancer by mechanisms that remain unclear. In silico analysis identified putative transcription factor specificity protein 1 (SP1) transcription factor binding sites in the miR-335 promoter. To investigate the relation between SP1 and miR-335, qRT-PCR was performed. Our results showed both Sp1 knockdown and mithramycin A increased miR-335 expression in ovarian cancer cell lines. Luciferase reporter assays indicated that Sp1 knockdown increased miR-335 transcriptional activity. ChIP experiments showed that Sp1 bound directly to miR-335 promoter. Moreover, transwell migration and wound-healing assays showed that Sp1 knockdown resulted in inhibited cell migration, which was in turn mitigated by miR-335 inhibitor. We propose that miR-335 was negatively regulated by SP1, which in turn contributes to miR-335 deregulation and tumor cells migration.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 23 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Guidenn Sulbarán, Antonio Biasutto, Franklin Méndez, Antonio Pinto, Lorenzo Alamo, Raúl Padrón〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Thick filaments from some striated muscles are regulated by phosphorylation of myosin regulatory light chains (RLCs). A tarantula thick filament quasi-atomic model achieved by cryo-electron microscopy has advanced our understanding on how this regulation occurs. In native thick filaments, an asymmetric intramolecular interaction between the actin-binding region of one myosin head (“blocked”) and the converter region of the other head (“free”) switches both heads off, establishing the myosin interacting-heads motif (IHM). This structural finding, together with motility assays, sequence analysis, and mass spectrometry (MS) observations have suggested a cooperative phosphorylation activation (CPA) mechanism for thick filament activation. In the CPA mechanism, some myosin free heads are phosphorylated constitutively in Ser35 by protein kinase C (PKC) and -under Ca〈sup〉2+〈/sup〉 control - others (free or blocked) heads temporally on Ser45 by myosin light chain kinase (MLCK), in a way that explains both force development and post-tetanic potentiation in tarantula striated muscle. We tested this model using MS to verify if Ca〈sup〉2+〈/sup〉-activation phosphorylates 〈em〉de novo〈/em〉 un-phosphorylated Ser35 heads. For this purpose, we standardized an approach based on 〈sup〉18〈/sup〉O isotopic ATP labeling to accurately detect by MS-MS the RLC phosphorylation under Ca〈sup〉2+〈/sup〉-activation. MS spectra showed 〈em〉de novo〈/em〉 〈sup〉18〈/sup〉O incorporation only on Ser45 but not on Ser35. As the constitutive Ser35 phosphorylation cannot be dephosphorylated, this result suggests that the number of RLCs on free heads with constitutively phosphorylated Ser35 does remain constant on Ca〈sup〉2+〈/sup〉-activation supporting that the myosin has a basal activation and force modulation or potentiation is controlled by MLCK Ser45 phosphorylation.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X20301078-fx1.jpg" width="423" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: Available online 23 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Santiago Loya-López, Alejandro Sandoval, Ricardo González-Ramírez, Aida Calderón-Rivera, Arturo Ávalos-Fuentes, Marina Rodríguez-Sánchez, René Caballero, Diana Tovar-Soto, Ricardo Felix, Benjamín Florán〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Neurotransmission is one of the most important processes in neuronal communication and depends largely on Ca〈sup〉2+〈/sup〉 entering synaptic terminals through voltage-gated Ca〈sup〉2+〈/sup〉 (Ca〈sub〉V〈/sub〉) channels. Although the contribution of L-type Ca〈sub〉V〈/sub〉 channels in neurotransmission has not been unambiguously established, increasing evidence suggests a role for these proteins in noradrenaline, dopamine, and GABA release. Here we report the regulation of L-type channels by Cdk5, and its possible effect on GABA release in the substantia nigra pars reticulata (SNpr). Using patch-clamp electrophysiology, we show that Cdk5 inhibition by Olomoucine significantly increases current density through Ca〈sub〉V〈/sub〉1.3 (L-type) channels heterologously expressed in HEK293 cells. Likewise, 〈em〉in vitro〈/em〉 phosphorylation showed that Cdk5 phosphorylates residue S1947 in the C-terminal region of the pore-forming subunit of Ca〈sub〉V〈/sub〉1.3 channels. Consistent with this, the mutation of serine into alanine (S1947A) prevented the regulation of Cdk5 on Ca〈sub〉V〈/sub〉1.3 channel activity. Our data also revealed that the inhibition of Cdk5 increased the frequency of high K〈sup〉+〈/sup〉-evoked miniature inhibitory postsynaptic currents in rat SNpr neurons, acting on L-type channels. These results unveil a novel regulatory mechanism of GABA release in the SNpr that involves a direct action of Cdk5 on L-type channels.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0006291X20301534-fx1.jpg" width="216" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: Available online 23 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Jie Liu, Yong He, Ke Cheng, Peng Xie〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Major depressive disorder (MDD) has become a potential cause of death and disability among young people worldwide. Numerous studies have indicated that the different cerebrospinal fluid (CSF) proteins may be used as mediums for MDD. Given the emergent interest of CSF proteins in MDD, we validated proteins expression in the choroid plexus (CP), the brain region that produces CSF in the lateral ventricle, the third ventricle, and the fourth ventricle of the central nervous system (CNS). The CSF constantly exchanges molecular substances with the brain tissue, which can dynamically reflect the metabolic microenvironment of the brain. In our previous study, Pepsin A (PGA) and periostin (POSTN) was associated with depressive-like behaviors of depressed macaca fascicularis models in CSF. Moreover, proteins that are expressed in the CP can be secreted into the CSF and may be associated MDD. This study sought to demonstrate the discrepancy of PGA and POSTN in the CP between Lipopolysaccharide (LPS)-induce depressed mice models and wild type (WT) mice. Our findings suggest that PGA and POSTN expression in CP of mice could be a possible candidate pathogenesis involved in MDD, which may contribute to a better understanding and treatment of MDD.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 23 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Biochemical and Biophysical Research Communications〈/p〉 〈p〉Author(s): Zhi Li, Pan-Pan Feng, Zhi-Bo Zhao, Wei Zhu, Jian-Ping Gong, Hui-Min Du〈/p〉