Identification and structural characterization of the 75-kDa rabbit zona pellucida protein

A cDNA (rc75) encoding a 75-kDa rabbit zona pellucida (ZP) glycoprotein (R75) has been cloned and sequenced. The predicted amino acid sequence consists of 676 amino acids including seven potential N-glycosylation sites. The cDNA hybridizes to a 2.4-kilobase mRNA in ovary that is not detectable in ot...

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Bibliographic Details
Published in:	The Journal of biological chemistry 1993-06, Vol.268 (17), p.12412-12417
Main Authors:	Lee, V H, Schwoebel, E, Prasad, S, Cheung, P, Timmons, T M, Cook, R, Dunbar, B S
Format:	Article
Language:	eng
Subjects:	Aging - metabolism Amino Acid Sequence Animals antigenic determinants Base Sequence Blotting, Northern cDNA Cloning, Molecular DNA Egg Proteins Female genes Membrane Glycoproteins - chemistry Membrane Glycoproteins - genetics Molecular Sequence Data Molecular Weight nucleotide sequence Ovary - growth & development Ovary - metabolism predictions Protein Structure, Secondary Rabbits Receptors, Cell Surface Recombinant Proteins - chemistry RNA - metabolism RNA, Messenger - analysis RNA, Messenger - metabolism Sequence Homology, Amino Acid Zona Pellucida - chemistry Zona Pellucida Glycoproteins zona pellucida protein
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Summary:	A cDNA (rc75) encoding a 75-kDa rabbit zona pellucida (ZP) glycoprotein (R75) has been cloned and sequenced. The predicted amino acid sequence consists of 676 amino acids including seven potential N-glycosylation sites. The cDNA hybridizes to a 2.4-kilobase mRNA in ovary that is not detectable in other rabbit tissues. The R75 mRNA was also found to be expressed during the early stages of rabbit ovarian development (2-6 weeks of age) when the ovary contains primordial, primary, and early secondary follicles. The deduced amino acid sequence of rc75 has 77% similarity to the mouse ZP2 protein but no similarity to mouse ZP3. R75 also contains regions with 35-55% similarity to a previously cloned rabbit 55-kDa ZP protein. Monte Carlo simulation comparison confirmed that R75 has a significant probability of homology with mouse ZP2 and the rabbit 55-kDa ZP protein. Antibodies were developed against a fragment of R75 (rc75a cDNA) expressed in the pEX expression vector. These antibodies were used to confirm that the expressed protein contained epitopes found in the native rabbit ZP glycoprotein. These data suggest that some, but not all, ZP proteins may be conserved among different species and that within a species ZP proteins may share similar regions.
ISSN:	0021-9258 1083-351X