Thurston R J, Korn N, Froman D P, Bodine A B
Department of Poultry Science, Clemson University, South Carolina 29634-0379.
Biol Reprod. 1993 Feb;48(2):393-402. doi: 10.1095/biolreprod48.2.393.
Protease and basic amidase activity was found in the seminal plasma of the domestic turkey. Amidase activity, measured through use of N-alpha-benzoyl-DL-arginine-p-nitroanilide-HCL (BAPNA), was 23-28 times greater for turkey than for guinea fowl or chicken. Within the reproductive tract, seminal plasma from the vas deferens had much greater activity than testicular or epididymal fluids. Turkey seminal plasma enzyme (TSPE) purified by chromatography or isoelectric focusing showed three protein bands by PAGE, each resolving on SDS-PAGE into two subunits with molecular weights of approximately 28,000-32,000 and 38,000-44,000. One of the three proteins also contained a larger subunit (M(r) 76,000-81,000) thought to be transferrin. Turkey acrosin consisted of three subunits with molecular weights below 20,500. Acrosin, but not TSPE, was visualized in native gels with N-alpha-benzoyl-DL-arginine-beta-naphthylamide (BANA)/Fast Garnet stain. Michaelis constants (BAPNA) for TSPE, acrosin, and trypsin were 2.41 +/- 0.12 x 10(-4) M (n = 5), 4.96-6.03 x 10(-4) M (n = 2), and 6.76 +/- 0.95 x 10(-4) M (n = 6), respectively. TSPE, like acrosin and trypsin, was inhibited by benzamidine but not iodoacetamide. While all natural trypsin inhibitors tested inhibited acrosin, TSPE was not inhibited by ovomucoid from chicken or turkey egg white.
在家养火鸡的精浆中发现了蛋白酶和碱性酰胺酶活性。通过使用N-α-苯甲酰-DL-精氨酸-对硝基苯胺盐酸盐(BAPNA)测定的酰胺酶活性,火鸡的比珍珠鸡或鸡高23 - 28倍。在生殖道内,输精管的精浆活性比睾丸或附睾液高得多。通过色谱法或等电聚焦纯化的火鸡精浆酶(TSPE)在聚丙烯酰胺凝胶电泳(PAGE)上显示出三条蛋白带,每条在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)上分解为两个亚基,分子量约为28,000 - 32,000和38,000 - 44,000。这三种蛋白质中的一种还包含一个较大的亚基(分子量76,000 - 81,000),被认为是转铁蛋白。火鸡顶体蛋白酶由分子量低于20,500的三个亚基组成。用N-α-苯甲酰-DL-精氨酸-β-萘基酰胺(BANA)/固红染色在天然凝胶中观察到顶体蛋白酶,但未观察到TSPE。TSPE、顶体蛋白酶和胰蛋白酶的米氏常数(BAPNA)分别为2.41±0.12×10⁻⁴M(n = 5)、4.96 - 6.03×10⁻⁴M(n = 2)和6.76±0.95×10⁻⁴M(n = 6)。TSPE与顶体蛋白酶和胰蛋白酶一样,被苯甲脒抑制,但不被碘乙酰胺抑制。虽然所有测试的天然胰蛋白酶抑制剂都能抑制顶体蛋白酶,但TSPE不受鸡或火鸡蛋清中的卵类粘蛋白抑制。