Miska W, Croseck H, Schill W B
Section Andrologie der Dermatologischen Klinik und Poliklinik der Universität München.
Biol Chem Hoppe Seyler. 1988 Jun;369(6):493-6. doi: 10.1515/bchm3.1988.369.1.493.
A rapid and highly efficient procedure for purification of kininase II from human seminal plasma is described. After ultra centrifugation, the enzyme was purified by gel filtration on Sepharose 6B CL and ion exchange chromatography followed by affinity chromatography of EDTA-inhibited enzyme on bradykinin-Sepharose. The enzyme was specifically inhibited by Captopril and BPP9a but not by phosphoramidon. PAGE in the presence of sodium dodecyl sulfate under reducing conditions resulted in two major protein bands with apparent molecular masses of about 55 kDa and 65 kDa and two faint protein bands at higher molecular masses. Antibodies raised against the major protein bands showed full cross reactivity with all four protein bands. The presented data indicate that kininase II consists of subunits.
本文描述了一种从人精浆中快速高效纯化激肽酶II的方法。超速离心后，通过Sepharose 6B CL凝胶过滤和离子交换色谱对酶进行纯化，随后将EDTA抑制的酶在缓激肽琼脂糖上进行亲和色谱。该酶被卡托普利和BPP9a特异性抑制，但不被磷酰胺脒抑制。在还原条件下十二烷基硫酸钠存在下的聚丙烯酰胺凝胶电泳产生两条主要蛋白带，表观分子量约为55 kDa和65 kDa，以及两条较高分子量的 faint蛋白带。针对主要蛋白带产生的抗体与所有四条蛋白带均显示出完全交叉反应性。所呈现的数据表明激肽酶II由亚基组成。
