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Effects of antibodies against tubulin on the movement of reactivated sea urchin sperm flagella

Asai, David J. and Brokaw, Charles J. (1980) Effects of antibodies against tubulin on the movement of reactivated sea urchin sperm flagella. Journal of Cell Biology, 87 (1). pp. 114-123. ISSN 0021-9525. doi:10.1083/jcb.87.1.114.

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Antibodies binding to sea urchin flagellar outer-doublet tubulin have been isolated from rabbit sera by tubulin-affinity chromatography employing electrophoretically purified tubulin as the immobilized substrate. This procedure provides "induced" antitubulin antibody from immune sera and "spontaneous" antitubulin antibody from preimmune sera. These antitubulins were characterized in terms of their specificity, ability to bind to sea urchin axonemes, and effects on the motility of reactivated spermatozoa. Induced antitubulin antibody specifically reduced the bend angle and symmetry of the movement of demembranated reactivated spermatozoa without affecting the beat frequency. At identical concentrations, spontaneous antitubulin had no effect on motility. Affinity-purified induced antitubulins from three other rabbits all gave specific bend-angle inhibition, whereas their corresponding spontaneous antitubulins had no effect on the flagellar movement. The effects of antitubulin on microtubule sliding were examined by observing the sliding disintegration of elastase-digested axonemes induced by MgATP2+-. Affinity-purified induced antitubulin antibody, in quantities sufficient to completely paralyze reactivated flagella, did not inhibit microtubule sliding. The amplitude-inhibiting effect of induced antitubulin on reactivated spermatozoa may be caused by action on a mechanism responsible for controlling flagellar bending rather than by interference with the active sliding process. This is the first report of an antitubulin antibody having an inhibitory activity on microtubule-associated movement.

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Additional Information:© 1980 by The Rockefeller University Press. Received for publication 3 March 1980, and in revised form 2 June 1980. We thank Tom Simonick for his able assistance with portions of this work, Prof. Ray Owen for the use of his immunology facilities and for his advice, and David L. Gard and Prof. Elias Lazarides for helping with the immunofluorescence experiments. We thank the U. S. Public Health Service for support provided by National Institutes of Health grant GM 18711.
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NIHGM 18711
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Record Number:CaltechAUTHORS:ASAjbc80
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:4434
Deposited By: Lindsay Cleary
Deposited On:23 Aug 2006
Last Modified:08 Nov 2021 20:17

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