Kolk A H, Samuel T
Biochim Biophys Acta. 1975 Jun 26;393(2):307-19. doi: 10.1016/0005-2795(75)90057-4.
Human spermatozoa were solubilized by treatment with urea/guanidine and fractionated by ion-exchange chromatography with Bio-Rex 70 resin. Four fractions were obtained. The acidic proteins pass unretarded, whereas the moderately basic and two strongly basic protein fractions are eluted by means of guanidine gradients. The main protein components of the strongly basic protein fractions have been isolated by gel filtration on Sephadex G-50. The purified proteins have been named human sperm protamine 1 and 2. They contain 47 and 51 aminoacid residues (mol. wt 6280 and 6840), of which 22 and 24 are arginine and 5 and 4 are half cystine residues, respectively. The electrophoretic mobility in urea/polyacrylamide gels is between that of calf thymus histone and salmon protamine. Human protamine 1 and 2 are both auto-antigens as has been detected by a quantitative immunofluorescence inhibition test.
用人精子经尿素/胍处理使其溶解，并用Bio-Rex 70树脂通过离子交换色谱法进行分级分离。得到了四个级分。酸性蛋白质不被保留而直接通过，而中等碱性和两个强碱性蛋白质级分则通过胍梯度洗脱。通过在Sephadex G-50上进行凝胶过滤，已分离出强碱性蛋白质级分的主要蛋白质成分。纯化后的蛋白质被命名为人类精蛋白1和2。它们分别含有47和51个氨基酸残基（分子量分别为6280和6840），其中分别有22个和24个精氨酸残基，5个和4个半胱氨酸残基。在尿素/聚丙烯酰胺凝胶中的电泳迁移率介于小牛胸腺组蛋白和鲑鱼精蛋白之间。通过定量免疫荧光抑制试验检测发现，人类精蛋白1和2均为自身抗原。
