Alpha-actinin and calmodulin interact with distinct sites on the arms of the clathrin trimer.
In the present study, the interaction of alpha-actinin and calmodulin with clathrin heavy chain are demonstrated using Western blot analysis and rotary shadowing electron microscopy. The results show that alpha-actinin and calmodulin bind the clathrin heavy chain. The interaction is specific and affected by calcium. However, the interaction of both proteins with the clathrin heavy chain is distinct; the proteins do not block each other's ability to bind, and they interact with different protein fragments of the clathrin heavy chain. Furthermore, using rotary shadowing the results show that alpha-actinin differentially affected the terminal region of the clathrin trimer. Whereas, the effects of calmodulin were most noticeably detected along the length of trimer arms. The possible existence of distinct binding sites on the arms of the clathrin trimer for these cytosolic proteins supports the contention that these cytosolic proteins play an important role in cellular trafficking.
Merisko
E M
EM
Anatomy Department, University of Kansas Medical Center, Kansas City 66103.
Welch
J K
JK
Chen
T Y
TY
Chen
M
M
D000185:Actinin / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D001667:Binding, Competitive; D002118:Calcium / Q000494:pharmacology; D002147:Calmodulin / Q000378:metabolism* / Q000648:ultrastructure; D002417:Cattle; D002966:Clathrin / Q000032:analysis* / Q000648:ultrastructure; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D013997:Time Factors
D002147:Calmodulin; D002966:Clathrin; D046911:Macromolecular Substances; D000185:Actinin; D002118:Calcium