The Journal of Cell Biology, Vol 101, 924-941, Copyright © 1985 by The Rockefeller University Press

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Structure of the Chlamydomonas agglutinin and related flagellar surface proteins in vitro and in situ

UW Goodenough, WS Adair, P Collin-Osdoby and JE Heuser

Using the quick-freeze, deep-etch technique, we compare the structure of the cane-shaped plus and minus sexual agglutinin molecules purified from gametes of Chlamydomonas reinhardi. We also describe the structure of three additional gamete-specific fibrillar molecules, called short canes, loops, and crescents, which are structurally related to the agglutinins. Four non-agglutinating mutant strains are found to produce the three latter fibrils but not canes, supporting our identification of the cane-shaped molecule as the agglutinin. The heads of the plus and minus canes are shown to differ in morphology. Moreover, two treatments that inactivate the plus agglutinin in vitro--thermolysin digestion and disulfide reduction/alkylation--bring about detectable structural changes only in the head domain of the cane, suggesting that the head may play an indispensible role in affecting gametic recognition/adhesion. We also present quick-freeze, deep-etch images of the flagellar surfaces of gametic, vegetative, and mutant cells of Chlamydomonas reinhardi. The gametic flagella are shown to carry the canes, short canes, loops, and crescents present in in vitro preparations. The cane and crescent proteins self-associate on the flagellar surface into stout fibers of uniform caliber, and they align along the longitudinal axis of the flagellum. The short canes and loops co-purify with flagella but, in the presence of mica, dissociate so that they lie to the sides of the flagella. The agglutinin canes of both mating types are oriented with their hooks at the membrane surface and their heads directed outward, where they are positioned to participate in the initial events of sexual agglutination.
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This article has been cited by other articles:

• Lee, J.-H., Waffenschmidt, S., Small, L., Goodenough, U. (2007). Between-Species Analysis of Short-Repeat Modules in Cell Wall and Sex-Related Hydroxyproline-Rich Glycoproteins of Chlamydomonas. Plant Physiol. 144: 1813-1826 [Abstract] [Full Text]  
• Ferris, P. J., Waffenschmidt, S., Umen, J. G., Lin, H., Lee, J.-H., Ishida, K., Kubo, T., Lau, J., Goodenough, U. W. (2005). Plus and Minus Sexual Agglutinins from Chlamydomonas reinhardtii. Plant Cell 17: 597-615 [Abstract] [Full Text]  

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