An antiserum against a glycoprotein functional in flagellar adhesion between Chlamydomonas eugametos gametes |
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Authors: | Peter F Lens Frans Olofsen Anneke Nederbragt Alan Musgrave Herman van den Ende |
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Institution: | (1) Department of Plant Physiology, University of Amsterdam, Kruislaan 318, Amsterdam, The Netherlands |
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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.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
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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Keywords: | Chlamydomonas Agglutination Flagella Glycoproteins Anti-PAS-1 2 serum |
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