Büllesbach E E, Schwabe C
Biochemistry. 1985 Dec 17;24(26):7717-22. doi: 10.1021/bi00347a032.
Porcine ovaries were collected from pregnant sows under conditions designed to keep autolysis to an absolute minimum. During the extraction the tissues were never allowed to warm up to 0 degree C until submerged in 1.6 N HCl. Isolation and fractionation of the various relaxin forms became possible by application of CM-cellulose chromatography at pH 5.5 and 7.8, gel filtration, and high-performance liquid chromatography. The new isolation procedure has made it possible to isolate and identify LeuB32 relaxin. Also, [Leu-PheA0]B29 relaxin was identified and the existence of a [Leu-PheA0]B32 relaxin may be deduced from our data. Controlled digestion of B-chain-extended relaxins with carboxypeptidase A led to the large-scale production of homogeneous B29 relaxin, a suitable starting material for controlled chemical modification of porcine relaxin.
从怀孕母猪体内采集猪卵巢,采集条件旨在将自溶现象控制在绝对最低限度。在提取过程中,组织在浸入1.6N盐酸之前,温度绝不能升至0摄氏度。通过在pH值5.5和7.8条件下应用CM-纤维素色谱法、凝胶过滤法和高效液相色谱法,各种松弛素形式的分离和分级成为可能。新的分离程序使得分离和鉴定LeuB32松弛素成为可能。此外,已鉴定出[Leu-PheA0]B29松弛素,并且从我们的数据中可以推断出[Leu-PheA0]B32松弛素的存在。用羧肽酶A对B链延长的松弛素进行可控消化,可大规模生产出均一的B29松弛素,这是对猪松弛素进行可控化学修饰的合适起始材料。