Siezen R J
Biophys Chem. 1984 Jan;19(1):49-55. doi: 10.1016/0301-4622(84)85005-x.
Crystallins from calf lens were subjected to ultrafiltration through an Amicon XM-300 membrane to determine whether specific interactions between identical proteins (self-association) or different proteins (mixed-association) could be detected and quantified. Single crystallins at different concentrations, simple mixtures and total lens extracts were studied separately. alpha-Crystallin (Mr 800 000) is nearly fully retained (greater than 95%) by XM-300. Retention of beta-crystallins (Mr 50 000-200 000) is found to be much higher than expected from their molecular weights. Ultrafiltration of gamma-crystallin (Mr 20 000) solutions of 1.0-22.6 g/l shows that retention increases as a function of protein concentration. In solutions of single crystallins, self-association effects could not be separated from concentration polarization effects at the membrane surface. In mixtures of crystallins, mixed-association could not be separated from self-association, concentration polarization and excluded volume effects on self-association.
将来自小牛晶状体的晶状体蛋白通过Amicon XM - 300膜进行超滤，以确定是否能够检测和定量相同蛋白质之间的特异性相互作用（自缔合）或不同蛋白质之间的特异性相互作用（混合缔合）。分别研究了不同浓度的单一晶状体蛋白、简单混合物和晶状体总提取物。α - 晶状体蛋白（分子量800000）几乎被XM - 300完全保留（大于95%）。发现β - 晶状体蛋白（分子量50000 - 200000）的保留率远高于根据其分子量预期的保留率。对浓度为1.0 - 22.6 g/l的γ - 晶状体蛋白（分子量20000）溶液进行超滤表明，保留率随蛋白质浓度的增加而增加。在单一晶状体蛋白溶液中，自缔合效应无法与膜表面的浓度极化效应区分开来。在晶状体蛋白混合物中，混合缔合无法与自缔合、浓度极化以及自缔合的排阻体积效应区分开来。
