The SH3 domain of amphiphysin binds the proline-rich domain of dynamin at a single site that defines a new SH3 binding consensus sequence.
Amphiphysin is an SH3 domain-containing neuronal protein that is highly concentrated in nerve terminals where it interacts via its SH3 domain with dynamin I, a GTPase implicated in synaptic vesicle endocytosis. We show here that the SH3 domain of amphiphysin, but not a mutant SH3 domain, bound with high affinity to a single site in the long proline-rich region of human dynamin I, that this site was distinct from the binding sites for other SH3 domains, and that the mutation of two adjacent amino acids in dynamin I was sufficient to abolish binding. The dynamin I sequence critically required for amphiphysin binding (PSRPNR) fits in the novel SH3 binding consensus identified for the SH3 domain of amphiphysin via a combinatorial peptide library approach: PXRPXR(H)R(H). Our data demonstrate that the long proline-rich stretch present in dynamin I contained multiple SH3 domain binding sites that recognize interacting proteins with high specificity.
Grabs
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Department of Cell Biology and Howard Hughes Medical Institute, Yale University School of Medicine, Boyer Center for Molecular Medicine, New Haven, Connecticut 06510, USA.
Slepnev
V I
VI
Songyang
Z
Z
David
C
C
Lynch
M
M
Cantley
L C
LC
De Camilli
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000378:metabolism; D034284:Dynamin I; D034281:Dynamins; D020558:GTP Phosphohydrolases / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011392:Proline / Q000378:metabolism; D011817:Rabbits; D017421:Sequence Analysis; D018909:src Homology Domains / Q000235:genetics
D009419:Nerve Tissue Proteins; C075637:amphiphysin; D011392:Proline; D034284:Dynamin I; D020558:GTP Phosphohydrolases; D034281:Dynamins