PURIFICATION OF A SEX-SPECIFIC LECTIN INVOLVED IN GAMETE BINDING OF AGLAOTHAMNION CALLOPHYLLIDICOLA (RHODOPHYTA)

Egg and sperm binding and correct recognition is the first stage for successful fertilization. In red algae, spermatial attachment to female trichogynes is mediated by a specific binding between the lectin(s) distributed on the surface of trichogyne and the complementary carbohydrates on the spermat...

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Bibliographic Details
Published in:Journal of phycology 2012-08, Vol.48 (4), p.916-924
Main Authors: Shim, Eunyoung, Shim, Junbo, Klochkova, Tatyana A., Han, Jong Won, Kim, Gwang Hoon
Format: Article
Language:English
Subjects:
Aglaothamnion
Aglaothamnion callophyllidicola
fertilization
gamete recognition
lectin
Rhodophyta
sex-specific
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Summary:Egg and sperm binding and correct recognition is the first stage for successful fertilization. In red algae, spermatial attachment to female trichogynes is mediated by a specific binding between the lectin(s) distributed on the surface of trichogyne and the complementary carbohydrates on the spermatial surface. A female-specific lectin was isolated from Aglaothamnion callophyllidicola by agarose-bound fetuin affinity chromatography. Two proteins, 50 and 14 kDa, eluted from the fetuin column were separated using a native-polyacrylamide gel electrophoresis method and subjected to a gamete binding assay. The 50 kDa protein, which blocked spermatial binding to female trichogynes, was used for further analysis. Internal amino acid sequence of the 50 kDa protein was analyzed using matrix-assisted laser desorption/ionization-mass spectrometry and degenerated primers were designed based on the information. A full-length cDNA encoding the lectin was obtained using rapid amplification of cDNA ends polymerase chain reaction (PCR). The cDNA was 1552 bp in length and coded for a protein of 450 amino acids with a deduced molecular mass of 50.7 kDa, which agreed well with the protein data. Real-time PCR analysis showed that this protein was up-regulated about 10-fold in female thalli. As the protein was novel and showed no significant homology to any known proteins, it was designated Rhodobindin.
ISSN:0022-3646
1529-8817
DOI:10.1111/j.1529-8817.2012.01155.x