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  • 1
    Publication Date: 2013-08-13
    Description: The internal organization of eukaryotic cells into functionally specialized, membrane-delimited organelles of unique composition implies a need for active, regulated lipid transport. Phosphatidylserine (PS), for example, is synthesized in the endoplasmic reticulum and then preferentially associates--through mechanisms not fully elucidated--with the inner leaflet of the plasma membrane. Lipids can travel via transport vesicles. Alternatively, several protein families known as lipid-transfer proteins (LTPs) can extract a variety of specific lipids from biological membranes and transport them, within a hydrophobic pocket, through aqueous phases. Here we report the development of an integrated approach that combines protein fractionation and lipidomics to characterize the LTP-lipid complexes formed in vivo. We applied the procedure to 13 LTPs in the yeast Saccharomyces cerevisiae: the six Sec14 homology (Sfh) proteins and the seven oxysterol-binding homology (Osh) proteins. We found that Osh6 and Osh7 have an unexpected specificity for PS. In vivo, they participate in PS homeostasis and the transport of this lipid to the plasma membrane. The structure of Osh6 bound to PS reveals unique features that are conserved among other metazoan oxysterol-binding proteins (OSBPs) and are required for PS recognition. Our findings represent the first direct evidence, to our knowledge, for the non-vesicular transfer of PS from its site of biosynthesis (the endoplasmic reticulum) to its site of biological activity (the plasma membrane). We describe a new subfamily of OSBPs, including human ORP5 and ORP10, that transfer PS and propose new mechanisms of action for a protein family that is involved in several human pathologies such as cancer, dyslipidaemia and metabolic syndrome.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Maeda, Kenji -- Anand, Kanchan -- Chiapparino, Antonella -- Kumar, Arun -- Poletto, Mattia -- Kaksonen, Marko -- Gavin, Anne-Claude -- England -- Nature. 2013 Sep 12;501(7466):257-61. doi: 10.1038/nature12430. Epub 2013 Aug 11.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉European Molecular Biology Laboratory, EMBL, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/23934110" target="_blank"〉PubMed〈/a〉
    Keywords: Biological Transport ; Carrier Proteins/chemistry/metabolism ; Cell Membrane/metabolism ; Dyslipidemias/metabolism ; Endoplasmic Reticulum/metabolism ; Homeostasis ; Humans ; Metabolic Syndrome X/metabolism ; Neoplasms/metabolism ; Phosphatidylserines/*metabolism ; Phylogeny ; *Protein Interaction Maps ; Receptors, Steroid/chemistry/*metabolism ; Saccharomyces cerevisiae/*metabolism ; Saccharomyces cerevisiae Proteins/chemistry/*metabolism ; Substrate Specificity
    Print ISSN: 0028-0836
    Electronic ISSN: 1476-4687
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
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  • 2
    Publication Date: 2015-06-20
    Description: During clathrin-mediated endocytosis (CME), plasma membrane regions are internalized to retrieve extracellular molecules and cell surface components. Whether endocytosis occurs by direct clathrin assembly into curved lattices on the budding vesicle or by initial recruitment to flat membranes and subsequent reshaping has been controversial. To distinguish between these models, we combined fluorescence microscopy and electron tomography to locate endocytic sites and to determine their coat and membrane shapes during invagination. The curvature of the clathrin coat increased, whereas the coated surface area remained nearly constant. Furthermore, clathrin rapidly exchanged at all stages of CME. Thus, coated vesicle budding appears to involve bending of a dynamic preassembled clathrin coat.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Avinoam, Ori -- Schorb, Martin -- Beese, Carsten J -- Briggs, John A G -- Kaksonen, Marko -- New York, N.Y. -- Science. 2015 Jun 19;348(6241):1369-72. doi: 10.1126/science.aaa9555.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Cell Biology and Biophysics Unit, The European Molecular Biology Laboratory, Heidelberg 69117, Germany. Structural and Computational Biology Unit, The European Molecular Biology Laboratory, Heidelberg 69117, Germany. ; Structural and Computational Biology Unit, The European Molecular Biology Laboratory, Heidelberg 69117, Germany. Electron Microscopy Core Facility, The European Molecular Biology Laboratory, Heidelberg 69117, Germany. ; Cell Biology and Biophysics Unit, The European Molecular Biology Laboratory, Heidelberg 69117, Germany. ; Structural and Computational Biology Unit, The European Molecular Biology Laboratory, Heidelberg 69117, Germany. Cell Biology and Biophysics Unit, The European Molecular Biology Laboratory, Heidelberg 69117, Germany. marko.kaksonen@unige.ch john.briggs@embl.de. ; Cell Biology and Biophysics Unit, The European Molecular Biology Laboratory, Heidelberg 69117, Germany. Structural and Computational Biology Unit, The European Molecular Biology Laboratory, Heidelberg 69117, Germany. marko.kaksonen@unige.ch john.briggs@embl.de.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/26089517" target="_blank"〉PubMed〈/a〉
    Keywords: Cell Line ; Clathrin/*chemistry ; Coated Pits, Cell-Membrane/*chemistry ; Electron Microscope Tomography ; *Endocytosis ; Fluorescence Recovery After Photobleaching ; Humans ; Microscopy, Fluorescence
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
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