Synaptotagmin regulation of coated pit assembly.
Synaptotagmins bind clathrin AP-2 with high affinity via their second C(2) domain, which indicates they are involved in coated pit function. We now report that expression of synaptotagmins lacking either the second C(2) domain or the entire cytoplasmic region potently inhibit endocytosis. Inhibition was dependent on two intramembrane cysteine residues that were found to be essential for synaptotagmin oligomerization. Cells expressing the wild-type, but not the mutant, truncated synaptotagmin fragment had a reduced number of clathrin-coated pits. These results suggest that the formation of synaptotagmin multimers is an important step in the regulation of coated pit assembly.
von Poser
C
C
Departments of Cell Biology, Biochemistry, and Molecular Genetics and The Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, Texas 75235, USA.
Zhang
J Z
JZ
Mineo
C
C
Ding
W
W
Ying
Y
Y
Sudhof
T C
TC
Anderson
R G
RG
D033965:Adaptor Protein Complex alpha Subunits; D033942:Adaptor Proteins, Vesicular Transport; D002135:Calcium-Binding Proteins; D002460:Cell Line; D003034:Coated Pits, Cell-Membrane / Q000378:metabolism* / Q000502:physiology / Q000648:ultrastructure; D003545:Cysteine / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004705:Endocytosis; D005455:Fluorescent Antibody Technique; D005799:Genes, Dominant; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008957:Models, Genetic; D016297:Mutagenesis, Site-Directed; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D010957:Plasmids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D050857:Synaptotagmins; D014162:Transfection
D033965:Adaptor Protein Complex alpha Subunits; D033942:Adaptor Proteins, Vesicular Transport; D002135:Calcium-Binding Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D016271:Proto-Oncogene Proteins c-myc; D011994:Recombinant Proteins; D050857:Synaptotagmins; D003545:Cysteine