Jonáková V, Kraus M, Veselský L, Cechová D, Bezouska K, Tichá M
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
J Reprod Fertil. 1998 Sep;114(1):25-34. doi: 10.1530/jrf.0.1140025.
Heparin-binding proteins (designated BHB-2-BHB-9) were isolated from boar seminal plasma by affinity chromatography on heparin immobilized on polyacrylamide gel, followed by reverse phase HPLC. According to their N-terminal amino acid sequences, BHB-3-BHB-5 belong to the AQN family of spermadhesins and BHB-7-BHB-9 to the AWN family. BHB-6 is composed of two different proteins. The dominant protein (14 kDa) has the N-terminal amino acid sequence HNKQEGRDHD that is identical to the sequence of human semenogelin at positions 85-94. The minor proteins (16 and 17 kDa) belong to the AWN family of spermadhesins. The 14 kDa HNK protein does not crossreact with antibodies against AQN or AWN spermadhesins. BHB-2 also binds to the acrosome of boar epididymal spermatozoa but has the N-terminal sequence DQH. Therefore, basic protein BHB-2 belongs to a new family of DQH sperm surface proteins that are homologous to the acidic proteins from bull and stallion seminal plasma, to the collagen binding domain II in fibronectin and to the leucocyte cell-cell adhesion regulator, but are not homologous to AQN or AWN spermadhesins. Nevertheless, anti-AQN-1 spermadhesin antibodies crossreact strongly with DQH protein. All boar heparin-binding proteins bind concanavalin A indicating their glycoprotein nature, which was proved by the detection of glucosamine and galactosamine residues in their molecules. Furthermore, spermadhesins interact with zona pellucida, protease inhibitors and a polyacrylamide derivative of heparin. Affinity chromatography experiments showed that the DQH protein bound to gelatin-agarose together with the AWN proteins and that the DQH protein and AQN-1 spermadhesin belong to the phosphoryl choline binding proteins.
通过在固定于聚丙烯酰胺凝胶上的肝素上进行亲和层析，随后进行反相高效液相色谱，从公猪精浆中分离出肝素结合蛋白（命名为BHB - 2 - BHB - 9）。根据它们的N端氨基酸序列，BHB - 3 - BHB - 5属于精子黏附素的AQN家族，BHB - 7 - BHB - 9属于AWN家族。BHB - 6由两种不同的蛋白质组成。主要蛋白质（14 kDa）的N端氨基酸序列为HNKQEGRDHD，与人类精液凝固蛋白在85 - 94位的序列相同。次要蛋白质（16和17 kDa）属于精子黏附素的AWN家族。14 kDa的HNK蛋白不与抗AQN或AWN精子黏附素的抗体发生交叉反应。BHB - 2也与公猪附睾精子的顶体结合，但其N端序列为DQH。因此，碱性蛋白BHB - 2属于一个新的DQH精子表面蛋白家族，该家族与公牛和种马精浆中的酸性蛋白、纤连蛋白中的胶原结合结构域II以及白细胞细胞间黏附调节剂同源，但与AQN或AWN精子黏附素不同源。然而，抗AQN - 1精子黏附素抗体与DQH蛋白强烈交叉反应。所有公猪肝素结合蛋白都能结合伴刀豆球蛋白A，表明它们具有糖蛋白性质，这通过在其分子中检测到氨基葡萄糖和半乳糖胺残基得到证实。此外，精子黏附素与透明带、蛋白酶抑制剂以及肝素的聚丙烯酰胺衍生物相互作用。亲和层析实验表明，DQH蛋白与AWN蛋白一起结合到明胶 - 琼脂糖上，并且DQH蛋白和AQN - 1精子黏附素属于磷酸胆碱结合蛋白。
