Purification and Molecular Cloning of Bovine Oviduct-Specific Glycoprotein1

Abstract

A specific 85-97-kDa (95-kDa) glycoprotein was found in bovine oviductal tissue and fluid during the follicular phase. In this study, a 95-kDa bovine oviductal glycoprotein (95-kDa BOGP) was purified by wheat germ agglutinin affinity and Mono-Q ion-exchange column chromatography. The first 29 NH2-terminal amino acid residues were determined by gas-phase microsequencing. A cDNA expression library prepared from poly(A)+ RNA isolated from bovine oviducts was screened with a monoclonal antibody to 95-kDa BOGP. A single positive clone containing a approximately 2-kb cDNA insert was isolated. The coding region contained 1612 bp translating to 537 amino acids. The derived amino acid sequence contained a partial signal sequence of 18 amino acids followed by 29 amino acids that were identical to the NH2-terminal amino acids determined by protein sequencing of purified 95-kDa BOGP. The amino acid sequence predicted a mature protein of 519 amino acids (57,684 daltons) containing one potential N-linked glycosylation site and five cysteines. Northern blot hybridization with a digoxigenin-labeled probe indicated that a single message of approximately 2.5 kb was present in oviductal RNA, and this message was detected in significantly greater amounts in oviductal RNA during the follicular phase than during the luteal phase. The amino acid sequence of a portion of 95-kDa BOGP was highly homologous (71% identity) to that of a baboon oviduct-specific glycoprotein.

Oviductins possess chitinase‐ and mucin‐like domains: A lead in the search for the biological function of these oviduct‐specific ZP‐associating glycoproteins

“…These characterized clones enclose only one similar unit of 15 amino acids but other related truncated elements are present. The number of potential sites of N-glycosylation ranges from one in the cow (Sendai et al, 1993) to seven in the hamster (Merlen et al, 19941, as deduced from their amino acid sequences (see Table 2). …”

“…Analyses of post-translational modifications using lectin binding experiments revealed that the mouse (Kapur and Johnson, 1985), bovine (Sendai et al, 1993), and hamster (Malette and Bleau, 1993) oviductins react with the WGA lectin thus indicating the presence of terminal a-D-NeuAc and/or nonterminal p-D-(GlcNAc)2 residues in their oligosaccharide side chains. Bovine oviductin binds peanut lectin (Wegner and Killian, 1992) which is specific for galactosyl (p1,3)Nacetylgalactosamine residues, confirming that it bears Sequence comparison of the repeated motifs found in oviductins.…”

Improving bovine semen diluents: insights from the male and female reproductive tracts, and the potential relevance of cervical mucins

“…Specific proteins such as osteopontin (OPN) (Gabler et al, 2003) and oviduct specific glycoprotein (OGP) (Boice et al, 1990;Sendai et al, 1994;Buhi, 2002) have been identified as secretory products in the bovine oviduct and were found to interact with gametes while also improving the efficiency of IVF (Martus et al, 1998;Killian, 2011). OPN is an extracellular matrix protein (McKee and Nanci, 1996) that, as well as being present in bovine seminal plasma (Cancel et al, 1997), is secreted by the oviduct mucosa.…”

“…OGPs (Boice et al, 1990;Sendai et al, 1994;Buhi, 2002) are present in the oviduct of every mammal studied to date. They are virtually exclusive to oviduct tissue and their production is greatest during oestrus and ovulation (Buhi, 2002).…”

Expression of Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) and Collagenase Genes in Preimplantation Bovine Embryos.

“…The restriction enzyme cleavage patterns of the bovine TIMP-1 fragments generated by Pst I and Sma I were analyzed to confirm the identity of the PCR products. In some experiments, the PCR fragments were electrophoresed on 2% agarose gels, capillary blotted onto Hybond N+ positively charged nylon membrane (Amersham, Buckinghamshire, UK), and hybridized with a DIGlabeled PCR probe prepared by the modified PCR method described previously [19].…”