Rat sperm galactosyl receptor: purification and identification by polyclonal antibodies raised against multiple antigen peptides

Molecular Reproduction and Development
J R MertzA L Kierszenbaum

Abstract

We have previously reported the purification of rats testis galactosyl receptor, an equivalent to the Ca(2+)-dependent (C-type) minor variant of rat hepatic lectin-2/3 (RHL-2/3). We now report the purification of galactosyl receptor from rat sperm and its immunolocalization in the intact rat testis and sperm by polyclonal antibodies prepared using multiple antigen peptides (MAP) as immunogens. Two MAP antigens (designated 27-mer and 28-mer), corresponding to amino acid sequences of the carbohydrate-recognition domain (galactose) and adjacent Ca(2+)-binding sites of RHL-2/3, were used for immunization. Anti-RHL-2/3, anti-p27, and anti-p28 sera crossreacted with rat hepatocyte RHL-2/3 and its rat testis and sperm equivalent, galactosyl receptor, purified by chromatofocusing followed by galactose-Hydropore-EP affinity chromatography. Neither anti-p27 nor anti-p28 sera cross-reacted with the major hepatocyte variant, RHL-1. A RHL-1-equivalent was not detected in rat testis and sperm. Immunofluorescence studies demonstrated that anti-p27 and anti-p28 sera recognize galactosyl receptor sites at the Sertoli cell-spermatogenic cell interface and on the dorsal surface of the sperm head, overlying the acrosome. The characteristic crescen...Continue Reading

References

Jul 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·J D Bleil, P M Wassarman
Aug 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·J P Tam
Oct 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·M Spiess, H F Lodish
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Jan 1, 1993·Annual Review of Cell Biology·K Drickamer, M E Taylor

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Citations

Sep 1, 1996·Molecular Reproduction and Development·L L TresA L Kierszenbaum
Jun 22, 2000·Molecular Reproduction and Development·E RivkinA L Kierszenbaum
Apr 4, 2003·Biology of Reproduction·P C N ChiuW S B Yeung
Nov 10, 2000·Developmental Biology·K MochidaA L Kierszenbaum
Aug 18, 2004·Journal of Lipid Research·Loredana QuadroVittorio Colantuoni
Mar 25, 2006·Molecular Reproduction and Development·Abraham L KierszenbaumLaura L Tres
Sep 18, 1998·Andrologia·E Töpfer-PetersenJ J Calvete

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