Goodman I, Unterman R D, Solomon S M, Hiatt R B
J Chromatogr. 1979 Oct 19;178(1):125-38. doi: 10.1016/s0021-9673(00)89703-4.
The protein binding stoichiometry of small molecules is here determined on a nanomole scale by a simplified procedure utilizing chromatography on thin layers of cross-linked dextran gels. New data are presented on the thin layer chromatographic properties of representative ligands, including a-amino acids, peptides, dyes and fluorigenic reagents, in relation to their molecular weights, polar characteristics, gel water regain values and denaturants, providing criteria for the general application of this method to studies of ligand binding with large as well as small molecules. By this procedure coherin peptides, A1 and B1--4, respectively, bind to coherin C in the molar ratio, 2:1, with a binding constant of about 10(5) M-1. Coherin C is believed to act as a carrier peptide.
本文采用一种简化方法，利用交联葡聚糖凝胶薄层色谱法，在纳摩尔尺度上测定小分子的蛋白质结合化学计量。给出了包括α-氨基酸、肽、染料和荧光试剂在内的代表性配体的薄层色谱特性的新数据，这些数据与它们的分子量、极性特征、凝胶水回吸值和变性剂有关，为该方法广泛应用于研究配体与大分子及小分子的结合提供了标准。通过该方法，粘着蛋白肽A1和B1-4分别以2:1的摩尔比与粘着蛋白C结合，结合常数约为10(5) M-1。粘着蛋白C被认为是一种载体肽。
