ATP-Dependent Conformational Changes of Dynein: Evidence for Changes in the Interaction of Dynein Heavy Chain with the Intermediate Chain 1
1995 年 117 巻 4 号 p. 903-907
詳細
1995 年 117 巻 4 号 p. 903-907
Conformational changes of the dynein β heavy chain/intermediate chain 1 (IC1) complex from outer arm dynein of sea urchin sperm flagella were examined by means of cross-linking experiments using a bifunctional cross-linker, dimethylsuberimidate. Cross-linking of the β/IC1 complex in the absence of ATP and vanadate (Vi) produced five cross-linked products. Immunoblotting of the products with anti-β chain and anti-IC1 antibodies revealed that all of them were cross-linked between β chain and IC1. Cross-linking of the complex in the presence of ATP and Vi produced four cross-linked products, but their electrophoretic mobilities were different from those of the cross-linked products obtained in the absence of ATP and Vi. Immunoblotting showed that only one cross-linked product was formed by cross-linking between β and IC1 and others were formed by intramolecular cross-linking of the β chain. Quantitative analysis indicated that cross-linking between β and IC1 decreased in the presence of ATP and Vi. These results suggest that conformational changes of the β heavy chain occur and the interaction between β chain and IC1 changes during ATP hydrolysis.
