Conformational changes of dynein: mapping and sequence analysis of ATP/Vanadate-dependent trypsin-sensitive sites on the outer arm dynein b heavy chain from sea urchin sperm flagella.
Conformational changes of dynein during ATP hydrolysis are demonstrated by the difference in the tryptic fragments of the dynein heavy chain between in the absence and presence of ATP and vanadate. Here tryptic sites in the presence of ATP and vanadate (Tav sites) have been mapped on the betaheavy chain of outer arm dynein from sea urchin sperm flagella. Tav sites are located not only near the central catalytic domain which includes four P-loops, but also near the carboxyl-terminal coiled-coil region. The Tav2 site is located in the most carboxyl-terminal region, which is nearly 850 amino acid residues apart from the the fourth P-loop (P4 site). The region from the most amino-terminal Tav site (Tav1 site) to the Tav2 site covers approximately 2,100 amino acid residues, which is almost half the whole betaheavy chain. Comparison of the sequences around the tryptic sites of the sea urchin b chain and those of the dynein heavy chains from other organisms reveals that the sequence around the Tav1 site is highly conserved in both cytoplasmic and axonemal dyneins but that around Tav2 sites is only conserved in axonemal dyneins, suggesting functional differences in the Tav2 region between the two subfamilies of dynein.
Inaba
K
K
Asamushi Marine Biological Station, Graduate School of Science, Tohoku University, Asamushi, Aomori, Aomori 039-3501, Japan. inaba@biology.tohoku.ac.jp
D000255:Adenosine Triphosphate / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D002696:Chlamydomonas / Q000737:chemistry; D004398:Dyneins / Q000737:chemistry* / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D008297:Male; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D012617:Sea Urchins / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013082:Sperm Tail / Q000737:chemistry* / Q000378:metabolism*; D014357:Trypsin / Q000737:chemistry; D014638:Vanadates / Q000737:chemistry
D014638:Vanadates; D000255:Adenosine Triphosphate; D014357:Trypsin; D004398:Dyneins