Structure and mass of mammalian respiratory ciliary outer arm 19S dynein.
Mammalian respiratory ciliary outer arm dyneins isolated as the major ATPase peak migrating at 19S on sucrose density gradients were examined by transmission electron microscopy of negatively stained samples and scanning transmission electron microscopy of unstained samples. The predominant discrete particle structure observed was composed of two globular heads apparently connected by amorphous or indistinct material. The heads were either circular or slightly elliptical of mean 13 +/- 1 X 10 +/- 2 nm dimensions. The mass of this structure averaged 1.22 +/- 0.34 million daltons with the individual globular heads averaging 310 +/- 77 kilodaltons (kD). Negative staining revealed that one or both of the globular heads often contained a central accumulation of stain measuring 2.5 +/- 1 nm across. A second type of structure, appearing with lesser frequency in the 19S fraction than in the unfractionated dynein preparation loaded onto the sucrose gradient, was a single globular head of 13 +/- 1 X 10 +/- 2 nm often with 2 +/- 1 nm centrally accumulated stain and with or without an appendage. This one-headed particle thus resembled one-half of the two-headed particle. Mass measurements were lower, however, for isolated, single globular heads, averaging 220 +/- 111 kD. A third type of particle observed was a ring-like structure with 4 +/- 1 nm centrally accumulated stain and without appendages. The ring structure was slightly larger in diameter, 14 +/- 1 nm, and had a greater peripheral accumulation of negative stain than either of the one- or two-headed particles, suggesting that it was not derived therefrom.(ABSTRACT TRUNCATED AT 250 WORDS)
Hastie
A T
AT
Department of Medicine, Thomas Jefferson University, Philadelphia, PA 19107.
Marchese-Ragona
S P
SP
Johnson
K A
KA
Wall
J S
JS
D000251:Adenosine Triphosphatases / Q000032:analysis; D000818:Animals; D002499:Centrifugation, Density Gradient; D002923:Cilia / Q000032:analysis; D004398:Dyneins / Q000648:ultrastructure*; D008854:Microscopy, Electron; D011506:Proteins / Q000032:analysis; D012137:Respiratory System / Q000032:analysis*
D011506:Proteins; D000251:Adenosine Triphosphatases; D004398:Dyneins