Strosberg A D, Margolies M N, Haber E
J Immunol. 1975 Nov;115(5):1422-4.
Rabbit light chain 3315, prepared from a homogeneous antipneumococcal antibody, was subjected to hydrolysis by pepsin without prior reduction and alkylation of the intrachain disulfide bonds. Gel filtration of the hydrolysate on Sephadex G-10, G-15, and G-25 and ion exchange chromatography on SP-Sephadex yielded several disulfide bridge peptides. These were fully reduced and alkulated and sequenced by Edman degradation. The peptides were located in the light chain sequence determined in independent studies from our laboratory. The half-cystine residues in this KB rabbit chain are located at positions 23, 80, 88, 134, 171, 194, and 214. The extra disulfide bridge extends between residues 80 and 171, thus joining the variable and constant domains. This is consistent with x-ray diffraction crystallographic studies showing that the corresponding residues in human light chains are separated by a distance compatible with disulfide bond formation.
兔轻链3315由一种同源抗肺炎球菌抗体制备而成，在未预先还原和烷基化链内二硫键的情况下，用胃蛋白酶进行水解。水解产物在Sephadex G - 10、G - 15和G - 25上进行凝胶过滤，并在SP - Sephadex上进行离子交换色谱，得到了几种二硫键桥连肽。这些肽被完全还原、烷基化，并通过埃德曼降解法进行测序。这些肽位于我们实验室独立研究确定的轻链序列中。这条KB兔链中的半胱氨酸残基位于第23、80、88、134、171、194和214位。额外的二硫键桥连在第80和171位残基之间延伸，从而连接可变区和恒定区。这与X射线衍射晶体学研究结果一致，该研究表明人轻链中的相应残基之间的距离与二硫键形成相匹配。
