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The Journal of Cell Biology, Vol 58, 618-629, Copyright © 1973 by Rockefeller University Press

ARTICLE

EFFECTS OF TRYPSIN DIGESTION ON FLAGELLAR STRUCTURES AND THEIR RELATIONSHIP TO MOTILITY



Keith E. Summers 1 and I. R. Gibbons 1

1 From the Department of Biochemistry and Biophysics and the Pacific Biomedical Research Center, University of Hawaii, Honolulu, Hawaii 96822.

Dr. Summers' present address is Department of Biological Sciences, Carnegie-Mellon University, Pittsburgh, Pennsylvania 15213.

Flagellar axonemes isolated from sea urchin sperm were digested with trypsin for various time periods. The course of digestion was monitored turbidimetrically and was found to take two different courses depending on the presence or absence of ATP in the digestion mixture. It was found that ATP induced active disintegration of the axonemes after slight digestion. Samples of the digested axonemes were examined with the electron microscope to determine the effects of trypsin digestion on the substructures of the axonemes. The rate at which trypsin sensitized the axonemes to ATP paralleled the rate at which it damaged the radial spokes and the nexin links, while the dynein arms were removed much more slowly. The results suggest that inactive dynein arms form cross bridges between the adjacent doublet tubules in digested axonemes, and that when activated by the addition of ATP, they induce an active shearing force between adjacent doublets. The radial spokes and the nexin links are not directly involved in the production of mechanical force, but they may participate in regulating the sliding between tubules to produce a propagated bending wave.

Submitted on March 5, 1973
Revised on May 7, 1973


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