Abstract
Chlamydomonas eugametos gametes agglutinate sexually by their flagellar surfaces. The agglutination factor on mating type minus (mt-) gametes is thought to be a glycoprotein named PAS-1.2. To test this idea, an antiserum was raised against purified PAS-1.2., which reacted with isolated PAS-1.2 (immunoprecipitation tests) and blocked the ability of isolated PAS-1.2 to induce sexual twitching in mt + gametes. When tested with living cells, the antiserum specifically agglutinated mt - gametes and induced a reaction resembling twitching. Mt + flagella were shown to bind the antiserum (indirect immunofluorescence) but much less than mt - gametes. Mt - gametes pretreated with Fab fragments of the antiserum were unable to reproduce sexually, while treated mt + gametes were unaffected. This effect presumably results from the ability of the serum to block mt - sexual agglutination, for mt - isoagglutinin was completely inactivated by the serum, while mt + isoagglutinin was unaffected. It is therefore argued that PAS-1.2 is the in vivo mt - agglutination factor. However it is shown that the antiserum was able to react in vitro not only with PAS-1.2 but with several other proteins in both mt - and mt + flagella.
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Abbreviations
- SDS:
-
sodium dodecyl sulphate
- PAS:
-
periodic acid-Schiff
- GTC:
-
guanidine thiocyanate
- mt +/- :
-
mating type plus or minus
- PBS:
-
phosphate buffer-saline
- Fab:
-
univalent antibody fragment
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The investigations were supported by the Foundation for Fundamental Biological Research (BION), which is subsidized by the Netherlands Organization for the Advancement of Pure Research (Z.W.O.)
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Lens, P.F., Olofsen, F., Nederbragt, A. et al. An antiserum against a glycoprotein functional in flagellar adhesion between Chlamydomonas eugametos gametes. Arch. Microbiol. 131, 241–246 (1982). https://doi.org/10.1007/BF00405886
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DOI: https://doi.org/10.1007/BF00405886