期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2009
卷号：106
期号：32
页码：13359-13364
DOI：10.1073/pnas.0906945106
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：The large GTPase dynamin plays a key role in clathrin-mediated endocytosis in animal cells, although its mechanism of action remains unclear. Dynamins 1, 2, and 3 contain a pleckstrin homology (PH) domain that binds phosphoinositides with a very low affinity (KD > 1 mM), and this interaction appears to be crucial for function. These observations prompted the suggestion that an array of PH domains drives multivalent binding of dynamin oligomers to phosphoinositide-containing membranes. Although in vitro experiments reported here are consistent with this hypothesis, we find that PH domain mutations that abolish dynamin function do not alter localization of the protein in transfected cells, indicating that the PH domain does not play a simple targeting role. An alternative possibility is suggested by the geometry of dynamin helices resolved by electron microscopy. Even with one phosphatidylinositol-4,5-bisphosphate [PtdIns(4,5)P2] molecule bound per PH domain, these dynamin assemblies will elevate the concentration of PtdIns(4,5)P2 at coated pit necks, and effectively cluster (or sequester) this phosphoinositide. In vitro fluorescence quenching studies using labeled phosphoinositides are consistent with dynamin-induced PtdIns(4,5)P2 clustering. We therefore propose that the ability of dynamin to alter the local distribution of PtdIns(4,5)P2 could be crucial for the role of this GTPase in promoting membrane scission during clathrin-mediated endocytosis. PtdIns(4,5)P2 clustering could promote vesicle scission through direct effects on membrane properties, or might play a role in dynamin's ability to regulate actin polymerization.
