The substructure of isolated and in situ outer dynein arms of sea urchin sperm flagella.
Outer-arm dynein from the sperm of the sea urchin S. purpuratus was adsorbed to mica flakes and visualized by the quick-freeze, deep-etch technique. Replicas reveal particles comprised of two globular heads joined by two irregularly shaped stems which make contact along their length. One head is pear-shaped (18.5 X 12.5 nm) and the other is spherical (14.5-nm diam). The stems are decorated by a complex of bead-like subunits. The same two-headed protein is found in the 21S dynein-1 fraction of sucrose gradients. The beta-heavy chain/intermediate chain 1 (beta/IC-1) dynein subfraction, produced by low-salt dialysis and zonal centrifugation of the high-salt-extracted dynein-1, contains only single-headed molecules with single stems. These heads are predominantly pear-shaped (18.5 X 12.5 nm). Since 21S dynein-1 contains two heavy chains (alpha and beta), and the beta/IC-1 subfraction is comprised of only the beta-heavy chain (Tang et al., 1982, J. Biol. Chem. 257: 508-515), we conclude that each head is formed by a heavy chain, that the pear-shaped head contains the beta-heavy chain, and that the spherical head contains the alpha-heavy chain. The in situ outer dynein arms of demembranated sperm were also studied by the quick-freeze, deep-etch method. When frozen in reactivation buffer devoid of ATP, each arm consists of a large globular head that attaches to the A-microtubule by distally skewed subunits and attaches to the B-microtubule by a slender stalk. In ATP, this head shifts its orientation such that it can be seen to be constructed from two globular domains. We offer possible correlates between the in situ and the in vitro images, and we compare the structure of sea-urchin dynein with dynein previously described from Chlamydomonas and Tetrahymena.
Sale
W S
WS
Goodenough
U W
UW
Heuser
J E
JE
D000251:Adenosine Triphosphatases / Q000032:analysis*; D000818:Animals; D004398:Dyneins / Q000032:analysis*; D005613:Freeze Etching; D008297:Male; D008854:Microscopy, Electron; D008870:Microtubules / Q000032:analysis; D010455:Peptides / Q000302:isolation & purification; D011487:Protein Conformation; D012617:Sea Urchins; D013082:Sperm Tail / Q000201:enzymology*; D013094:Spermatozoa / Q000201:enzymology*
D010455:Peptides; D000251:Adenosine Triphosphatases; D004398:Dyneins