title: Interaction of vinculin with the clathrin heavy chain.
abstract: To further document the interaction of vinculin with the clathrin heavy chain (CHC) which was observed by using gel overlay, co-sedimentation experiments were performed and attempts were made to localize the domains involved on both molecules. The binding properties of proteolytic fragments of vinculin were investigated after cleavage with V8 protease. Neither the isolated globular domain, nor the C-terminal rod domain were able to interact with the CHC. Either the interaction involved the portion of vinculin which links these two domains, or the region of vinculin mediating the interaction was present on one of the two major fragments, but the cleavage itself resulted in conformational changes which abolished the binding. The first hypothesis could be ruled out using alpha-chymotrypsin generated fragments of vinculin, suggesting that the native conformation of vinculin might play an important role. Proteolytic cleavage of CHC with trypsin demonstrated that the interaction with vinculin is mediated by the proximal or distal segment of the CHC. Presence of clathrin light chain (CLC) associated with the CHC did not affect its interaction with vinculin. Vinculin did not interact with the CLC.
authors.affiliation: Institut de Biologie Médicale, Faculté de Médecine, Strasbourg, France.
authors.forename: J L
authors.initials: JL
authors.lastname: Fausser
authors.forename: E
authors.initials: E
authors.lastname: Ungewickell
authors.forename: J V
authors.initials: JV
authors.lastname: Ruch
authors.forename: H
authors.initials: H
authors.lastname: Lesot
mesh_terms: D000818:Animals; D002645:Chickens; D002918:Chymotrypsin / Q000378:metabolism; D002966:Clathrin / Q000737:chemistry / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005895:Gizzard, Avian / Q000737:chemistry; D015151:Immunoblotting; D010446:Peptide Fragments / Q000378:metabolism; D011487:Protein Conformation; D012697:Serine Endopeptidases / Q000378:metabolism; D016608:Talin / Q000378:metabolism; D016596:Vinculin / Q000737:chemistry / Q000378:metabolism*
chemical_list: D002966:Clathrin; D010446:Peptide Fragments; D016608:Talin; D016596:Vinculin; D012697:Serine Endopeptidases; D002918:Chymotrypsin; C028344:glutamyl endopeptidase