Handley C J, Bateman J F, Oakes B W, Lowther D A
Biochim Biophys Acta. 1975 Apr 29;386(2):444-50. doi: 10.1016/0005-2795(75)90287-1.
Cartilage cells from embryonic chick cartilage were grown in primary cultures. The cell layer was sequentially extracted with neutral saline, mercaptoethylamine and pepsin which revealed that these cells produced salt-soluble and salt-insoluble collagen. The alpha1- to alpha2-chain ratio was determined for the collagen extracted from the cultured cells and was found to be 13 to 1. Further analysis of the molecule was carried out by CNBr cleavage of the salt-extracted collagen and separation of resulting peptides by ion-exchange chromatography. It was shown that the cultured cartilage cells synthesize collagen of the type (alpha1[II])3.
取自胚胎期小鸡软骨的软骨细胞进行原代培养。细胞层依次用中性盐水、巯基乙胺和胃蛋白酶提取,结果表明这些细胞能产生盐溶性和盐不溶性胶原蛋白。测定了从培养细胞中提取的胶原蛋白的α1链与α2链的比例,发现为13比1。通过对盐提取的胶原蛋白进行溴化氰裂解并用离子交换色谱法分离所得肽段,对该分子进行了进一步分析。结果表明,培养的软骨细胞合成的是(α1[II])3型胶原蛋白。