Conformational changes of the beta chain of the outer-arm dynein from sea urchin sperm flagella coupled with ATP hydrolysis.
Conformational changes of the beta chain of the outer-arm dynein from sea urchin sperm flagella in relation to ATP hydrolysis was examined by tryptic digestion. Tryptic digestion of the beta chain in the presence of 2 mM ATP (ADP) and 100 microM vanadate (Vi) or in the presence of 4 mM ATP gamma S produced different polypeptides from in the case of no addition. The difference was similar to the result previously reported for 21S outer-arm dynein heavy chains [Inaba, K. & Mohri, H. (1989) J. Biol. Chem. 264, 8384-8388]. Unlike the tryptic digestion pattern of 21S dynein heavy chains, however, the 135-kDa polypeptide was consistently produced from the beta chain, even in the presence of ATP (ADP) and Vi. The tryptic digestion pattern of the 21S particle reconstituted from the separated a chain, the beta/IC1 complex and the IC2/IC3 complex [Tang, W.-J.Y., Bell, C.W., Sale, W.S., & Gibbons, I.R. (1982) J. Biol. Chem. 257, 508-515] was similar to that of intact 21S dynein; the 135-kDa polypeptide was only slightly produced in the presence of ATP and Vi. The digestion rate constant of the 135-kDa polypeptide from the beta chain in the presence of ATP and Vi was significantly decreased as compared with in the case of 21S dynein or that of the reconstituted 21S particle. These results suggest that the trypsin sensitivity of the 135-kDa region of the beta chain changes with the association of the beta/ICI complex with the alpha chain and the IC2/IC3 complex in the presence of ATP and Vi.
Inaba
K
K
Department of Biology, College of Arts and Sciences, University of Tokyo.
Ono
M
M
Mohri
H
H
D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D004398:Dyneins / Q000737:chemistry*; D006868:Hydrolysis; D008297:Male; D011487:Protein Conformation; D012617:Sea Urchins; D013082:Sperm Tail / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D014357:Trypsin / Q000494:pharmacology; D014638:Vanadates / Q000494:pharmacology
D014638:Vanadates; D000255:Adenosine Triphosphate; D014357:Trypsin; D004398:Dyneins