ALBERT

Beschreibung: Primitive basalts are rarely found in arcs. The active NW Rota-1 volcano in the Mariana arc has erupted near-primitive lavas, which we have sampled with ROV Hyper-Dolphin (HPD). Samples from the summit (HPD480) and eastern flank (HPD488) include 17 magnesian basalts (51–52 wt % SiO 2 ) with 7·5–9·5 wt % MgO and Mg-number of 61–67, indicating little fractionation. Olivine phenocrysts are as magnesian as Fo 93 and contain 0·4 wt % NiO; the Cr/(Cr + Al) values of spinels are mostly 0·5–0·8, indicating equilibrium with depleted mantle. There are three petrographic groups, based on phenocryst populations: (1) cpx–olivine basalt (COB); (2) plagioclase–olivine basalt (POB); (3) porphyritic basalt. Zr/Y and Nb/Yb are higher in POB (3·1–3·2 and 1·2–1·5, respectively) than in COB (Zr/Y = 2·8–3·0 and Nb/Yb = 0·7–0·9), suggesting that POB formed from lower degrees of mantle melting, or that the COB mantle source was more depleted. On the other hand, COB have Ba/Nb (70–80) and Th/Nb (0·4–0·5) that are higher than for POB (Ba/Nb = 30–35 and Th/Nb = 0·1–0·2), and also have steeper light rare earth element (LREE)-enriched patterns. Moreover, COB have enriched 87 Sr/ 86 Sr and 143 Nd/ 144 Nd, and higher Pb isotope values, suggesting that COB has a greater subduction component than POB. 176 Hf/ 177 Hf between COB and POB are similar and Hf behavior in COB and POB is similar to that of Zr, Y and HREE, suggesting that Hf is not included in the subduction component, which produced the differences between COB and POB. The calculated primary basaltic magmas of NW Rota-1 volcano (primary COB and POB magmas) indicate segregation pressures of 2–1·5 GPa (equivalent to 65–50 km depth). These magmas formed by 24–18% melting of mantle peridotite having Mg-number ~89·5. Diapiric ascent of hydrous peridotite mixed heterogeneously with sediment melts may be responsible for the NW Rota-1 basalts. These two basalt magma types are similar to those found at Sumisu and Torishima volcanoes in the Izu–Bonin arc, with COB representing wetter and POB representing drier magmas, where subduction zone-derived melt components are coupled with the water contents.

Beschreibung: Abnormalities in Z-disc proteins cause hypertrophic (HCM), dilated (DCM) and/or restrictive cardiomyopathy (RCM), but disease-causing mechanisms are not fully understood. Myopalladin (MYPN) is a Z-disc protein expressed in striated muscle and functions as a structural, signaling and gene expression regulating molecule in response to muscle stress. MYPN was genetically screened in 900 patients with HCM, DCM and RCM, and disease-causing mechanisms were investigated using comparative immunohistochemical analysis of the patient myocardium and neonatal rat cardiomyocytes expressing mutant MYPN. Cardiac-restricted transgenic (Tg) mice were generated and protein–protein interactions were evaluated. Two nonsense and 13 missense MYPN variants were identified in subjects with DCM, HCM and RCM with the average cardiomyopathy prevalence of 1.66%. Functional studies were performed on two variants (Q529X and Y20C) associated with variable clinical phenotypes. Humans carrying the Y20C-MYPN variant developed HCM or DCM, whereas Q529X-MYPN was found in familial RCM. Disturbed myofibrillogenesis with disruption of α-actinin2, desmin and cardiac ankyrin repeat protein (CARP) was evident in rat cardiomyocytes expressing MYPN Q529X . Cardiac-restricted MYPN Y20C Tg mice developed HCM and disrupted intercalated discs, with disturbed expression of desmin, desmoplakin, connexin43 and vinculin being evident. Failed nuclear translocation and reduced binding of Y20C-MYPN to CARP were demonstrated using in vitro and in vivo systems. MYPN mutations cause various forms of cardiomyopathy via different protein–protein interactions. Q529X-MYPN causes RCM via disturbed myofibrillogenesis, whereas Y20C-MYPN perturbs MYPN nuclear shuttling and leads to abnormal assembly of terminal Z-disc within the cardiac transitional junction and intercalated disc.

Beschreibung: The Myxococcus xanthus protein phosphatase Pph3 belongs to the Mg 2+ - or Mn 2+ -dependent protein phosphatase (PPM) family. Bacterial PPMs contain three divalent metal ions and a flap subdomain. Putative metal- or phosphate-ion binding site-specific mutations drastically reduced enzymatic activity. Pph3 contains a cyclic nucleotide monophosphate (cNMP)-binding domain in the C-terminal region, and it requires 2-mercaptoethanol for phosphatase activity; however, the C-terminal deletion mutant showed high activity in the absence of 2-mercaptoethanol. The phosphatase activity of the wild-type enzyme was higher in the presence of cAMP than in the absence of cAMP, whereas a triple mutant of the cNMP-binding domain showed slightly lower activities than those of wild-type, without addition of cAMP. In addition, mutational disruption of a disulphide bond in the wild-type enzyme increased the phosphatase activity in the absence of 2-mercaptoethanol, but not in the C-terminal deletion mutant. These results suggested that the presence of the C-terminal region may lead to the formation of the disulphide bond in the catalytic domain, and that disulphide bond cleavage of Pph3 by 2-mercaptoethanol may occur more easily with cAMP bound than with no cAMP bound.

Beschreibung: A reactive flow geochemical model based on pMELTS thermodynamic calculations explains the observed modal, major, and trace element variations in the Red Hills peridotite, New Zealand. The model also reproduces the major and trace element chemical variations in mid-ocean ridge basalts (MORB) observed in present-day spreading ridges. The Red Hills peridotite is thought to originate from palaeo-MOR magmatic processes in the mantle–Moho transition zone. The peridotite body consists of a harzburgite matrix and dunite channels. The harzburgite forms the Lower Unit and is intruded by replacive dunite channels in the Upper Unit. This lithology gradually turns into a massive dunite zone in which disseminated to lenticular clinopyroxene aggregates are present. The rare earth element (REE) abundances in the peridotite samples vary greatly depending on their lithologies. In the Lower Unit, REE are extremely depleted, whereas in the Upper Unit they are relatively enriched, in contradiction to the depleted lithologies. Our model consists of two stages. The first stage assumes melting of depleted MORB source mantle in the garnet stability field, and the second assumes reactions between residual solids and the melts from the first stage in the spinel stability field in an open system. The model explains the formation of depleted harzburgite and the formation of dunite channels in the harzburgite matrix well. The major and trace element compositions of the melts calculated by the model vary from ultra-depleted MOR melts in harzburgite to normal MORB in dunite, suggesting that these lithologies are residues of a palaeo-MOR. The model also explains the origins of the local and global geochemical trends observed in MORB and the geochemical variation in abyssal peridotite samples. Our model confirms the important role of reactive flow in the mantle–Moho transition zone beneath MORs.

Beschreibung: Reactive oxygen species (ROS) produced by plant NADPH oxidases (NOXes) are important in plant innate immunity. The Oryza sativa respiratory burst oxidase homologue B ( OsRbohB ) gene encodes a NOX the regulatory mechanisms of which are largely unknown. Here, we used a heterologous expression system to demonstrate that OsRbohB shows ROS-producing activity. Treatment with ionomycin, a Ca 2+ ionophore, and calyculin A, a protein phosphatase inhibitor, activated ROS-producing activity; it was thus OsRbohB activated by both Ca 2+ and protein phosphorylation. Mutation analyses revealed that not only the first EF-hand motif but also the upstream amino-terminal region were necessary for Ca 2+ -dependent activation, while these regions are not required for phosphorylation-induced ROS production.

Beschreibung: Proinsulin C-peptide shows beneficial effects on microvascular complications of Type 1 diabetes. However, the possible occurrence of membrane C-peptide receptor(s) has not been elucidated. The aim of this study was to identify and characterize membrane proteins to which C-peptide binds. The enzyme α-enolase was co-immunoprecipitated with C-peptide after chemical cross-linking to HL-60 cell surface proteins and identified by mass spectrometry. Recombinant α-enolase activity was modulated by C-peptide, with a significant decrease in K m for 2-phosphoglycerate without affecting V max . The enzyme modulation by C-peptide was abolished when C-terminal basic lysine residue (K434) of the enzyme was replaced by neutral alanine or acidic glutamate, but not with basic arginine. The enzyme modulation by C-peptide was reproduced with the C-peptide fragments containing glutamate corresponding to position 27 (E27) of the full-length C-peptide. Addition of a lysine analogue to the assay and A31 cell culture abrogated the enzyme modulation and MAP kinase activation by C-peptide, respectively. The results indicate that C-peptide has the capacity to activate α-enolase through a specific interaction between E27 of the peptide and K434 of the enzyme. Since α-enolase plays a role as a cell surface receptor for plasminogen, it may conceivably also serve as a receptor for C-peptide in vivo .

Beschreibung: Ferredoxin (Fd), which plays a pivotal role in photosynthesis as an electron carrier, forms a transient complex with various Fd-dependent enzymes, such as nitrite reductase (NiR), to achieve efficient intermolecular electron transfer. We studied the protein–protein interaction of Fd and NiR by NMR spectroscopy and determined three acidic regions of Fd to be major sites for the interaction with NiR, indicating that the complex is stabilized through electrostatic interaction. During this study, we found Fds from higher plant and cyanobacterium, in spite of their high structural similarities including the above acidic regions, differ remarkably in the interaction with cyanobacterial NiR. In activity assay of NiR, K m value for maize Fd (74.6 µM) was 9.6 times larger than that for Leptolyngbya boryana Fd (7.8 µM). The two Fds also showed a similar difference in binding assay to NiR-immobilized resin. Comparative site-specific mutagenesis of two Fds revealed that their discriminative ability for the interaction with NiR is attributed mainly to non-charged residues in the peripheral region of [2Fe–2S] cluster. These non-charged residues are conserved separately between Fds of plant and cyanobacterial origins. Our data highlight that intermolecular force(s) other than electrostatic attraction is(are) also crucial for the molecular interaction between Fd and partner enzyme.

Beschreibung: Modifications of rRNAs are clustered in functional regions of the ribosome. In Helix 74 of Escherichia coli 23S rRNA, guanosines at positions 2069 and 2445 are modified to 7-methylguanosine(m 7 G) and N 2 -methylguanosine(m 2 G), respectively. We searched for the gene responsible for m 7 G2069 formation, and identified rlmL , which encodes the methyltransferase for m 2 G2445, as responsible for the biogenesis of m 7 G2069. In vitro methylation of rRNA revealed that rlmL encodes a fused methyltransferase responsible for forming both m 7 G2069 and m 2 G2445. We renamed the gene rlmKL . The N-terminal RlmL activity for m 2 G2445 formation was significantly enhanced by the C-terminal RlmK. Moreover, RlmKL had an unwinding activity of Helix 74, facilitating cooperative methylations of m 7 G2069 and m 2 G2445 during biogenesis of 50S subunit. In fact, we observed that RlmKL was involved in the efficient assembly of 50S subunit in a mutant strain lacking an RNA helicase deaD .

Beschreibung: We present major element, trace element, and petrographic data on alkali basalts from St. Helena, and examine the geochemical characteristics of a recycled component involved in the source of HIMU ( 206 Pb/ 204 Pb 〉20·5) ocean island basalts. Petrographic and compositional variations in the St. Helena basalts are best explained by the combined effect of fractional crystallization and accumulation of phenocrysts. Primary melt compositions are estimated by correcting for the effects of crystal–liquid differentiation by reconstructing the order of crystallization and the relative amount of fractionated phases. This calculation indicates that the St. Helena alkali basalts are derived from a common primary magma with 14–20 wt % MgO. Simple partial melting of fertile mantle peridotite, depleted mid-ocean ridge basalt (MORB)-source mantle, or garnet pyroxenite fails to produce the St. Helena primary melt. Instead, this primary melt can be reproduced if there are contributions from ancient recycled oceanic crust and depleted peridotite [(Rb/Nb) PM = 0·38–0·80]. Subducted sediment can be excluded to explain the low (Rb, Ba, U)/Nb and Ce/Pb of St. Helena basalts. Geochemical modeling using major and trace element abundances, together with Sr, Nd, Pb, and Hf isotope ratios, indicates that the St. Helena primary melt can be formed by 1–2% melting of a peridotitic source that was refertilized by a small amount (8–18%) of melt derived from recycled oceanic crust. This source has a similar trace element pattern to modern normal (N)-MORB, but element abundances are 0·1–0·2 times N-MORB values. The calculated recycled crust has a wide range of present-day Pb isotopic ratios ( 206 Pb/ 204 Pb of 21·7–79·3 and 208 Pb/ 204 Pb of 40·8–89·3), 87 Sr/ 86 Sr of 0·7018–0·7028, 143 Nd/ 144 Nd of 0·51274–0·51285, and 176 Hf/ 177 Hf of 0·28262–0·28293 after a residence time of 1·2–2·8 Gyr. Rb, Ba, Pb, Sr, and light rare earth element abundances in the recycled crust are depleted compared with modern N-MORB, whereas Th, U, Sm, and Nd abundances fall within the range of compositional variations in modern N-MORB. The trace element compositions of the recycled oceanic crust can be explained by element behavior during seafloor alteration and subduction zone dehydration of oceanic crust. Therefore, recycling of ancient subducted oceanic crust is a potential process for producing the St. Helena HIMU basalts.