Adam M, Deyl Z
Ann Rheum Dis. 1984 Apr;43(2):258-63. doi: 10.1136/ard.43.2.258.
Collagen has been isolated and purified from the pronase digest of the outer annulus fibrosus of degenerated and unaffected intervertebral discs obtained at necropsy. The mixture of different collagen types has been purified and fractionated by 2 subsequent DEAE-cellulose chromatographic procedures according to established methods. Collagen chain polymers were then isolated by molecular sieving. Their reduction with 2-mercaptoethanol followed by agarose chromatography resulted in the separation of 3 major peaks corresponding to gamma, beta, and alpha chains. Cyanogen bromide (CNBr) peptides and aminoacid analyses revealed the identity of the isolated alpha-chain as alpha 1(III). Further support for this finding was obtained by immunofluorescence. It is suggested that the presence of collagen type III in the outer annulus fibrosus may be related to intervertebral disc prolapse.
已从尸检时获得的退变和未受影响的椎间盘外周纤维环的链霉蛋白酶消化物中分离并纯化了胶原蛋白。根据既定方法，通过2次连续的DEAE-纤维素色谱程序对不同类型胶原蛋白的混合物进行了纯化和分级分离。然后通过分子筛分离出胶原蛋白链聚合物。用2-巯基乙醇还原后再进行琼脂糖色谱分析，结果分离出了对应于γ、β和α链的3个主要峰。溴化氰（CNBr）肽和氨基酸分析表明，分离出的α链为α1(III)。免疫荧光进一步支持了这一发现。提示外周纤维环中III型胶原蛋白的存在可能与椎间盘突出有关。
