This study aimed to evaluate rabbit polyclonal antibodies raised against purified seminal plasma sperm membrance extracts (SP) heparin binding protein (HBP) for identifying HBPs in buffalo bull spermatozoa by western blotting. Anti-SP-HBP recognized 11 polypeptides in SDS-sperm membrance extracts (SME) of 31 tested bulls. Thirty one bulls were divided into G-1 (>40%) and G-II (≤40%) based on acrosome reaction. Immunoblotting revealed that HBPs of 24, 30, 38 and 43 kDa were present in 3%, 7.02%, 1.16% and 4.83% more bulls of G-I, whereas, 20 and 46 kDa HBPs were present in 13.2 and 9.65% more bulls of G-II. Immunoblotting of anti-HBP with sperm extracts of 10 bulls (22-31) indicated that 31 kDa positive bulls had 10.9% higher conception rate than 31 kDa negative bulls. Although 24 kDa HBP was detected in 10 bulls, but its expression was very weak in bull number 22, 23 and 26, which had 10.7% lower conception rate than the bulls with strong expression of 24 kDa HBP. In the present study, 17/20 kDa positive bulls exhibited 4.46% and 8.67% low conception rate than 17/20 kDa negative bulls. Mass spectrometry analysis revealed matching of 24, 31, 33 and 38 kDa proteins with MHC class 1 antigen, tRNA methyl transferase 11 homolog partial, parvalbumin alpha-like and cilia- and flagella-associated protein 99. This study suggests that buffalo bull fertility can be predicted from sperm HBP.

中文翻译：

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使用针对精浆血浆HBP的多克隆抗体鉴定精子肝素结合蛋白（HBP）：在水牛公牛育性中的应用

这项研究旨在评估针对纯化的精浆精子膜提取物（SP）肝素结合蛋白（HBP）产生的兔多克隆抗体，以通过Western印迹鉴定水牛牛精子中的HBP。抗SP-HBP识别了31只测试公牛的SDS-精子膜提取物（SME）中的11种多肽。根据顶体反应，将31头公牛分为G-1（> 40％）和G-II（≤40％）。免疫印迹显示，GI的公牛多存在3％，7.02％，1.16％和4.83％的HBP，分别为24、30、38和43 kDa，而G的公牛多存在20和46 kDa的HBP多于GI -II。用10头公牛（22-31）的精子提取物进行的抗HBP免疫印迹试验表明，31 kDa阳性公牛的受孕率比31 kDa阴性公牛高10.9％。尽管在10头公牛中检测到24 kDa HBP，但是在22号，23号和26号公牛中它的表达非常弱，其受孕率比24 kDa HBP强烈表达的公牛低。在本研究中，17/20 kDa阳性公牛比17/20 kDa阴性公牛的受孕率低4.46％和8.67％。质谱分析显示24、31、33和38 kDa蛋白与MHC 1类抗原，tRNA甲基转移酶11同源部分蛋白，小白蛋白α样蛋白以及纤毛和鞭毛相关蛋白99匹配。这项研究表明水牛的繁殖能力从精子HBP预测。