Kadler K E, Torre-Blanco A, Adachi E, Vogel B E, Hojima Y, Prockop D J
Department of Biochemistry and Molecular Biology, Jefferson Institute of Molecular Medicine, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 19107-6799.
Biochemistry. 1991 May 21;30(20):5081-8. doi: 10.1021/bi00234a035.
Type I procollagen was purified from cultured fibroblasts of a proband with a lethal variant of osteogenesis imperfecta. The protein was a mixture of normal procollagen and mutated procollagens containing a substitution of cysteine for glycine in either one pro alpha 1(I) chain or both pro alpha 1(I) chains, some or all of which were disulfide-linked through the cysteine at position alpha 1-748. The procollagen was then examined in a system for generating collagen fibrils de novo by cleavage of the pCcollagen to collagen with procollagen C-proteinase [Kadler et al. (1987) J. Biol. Chem. 262, 15696-15701]. The mutated collagens and normal collagens were found to form copolymers under a variety of experimental conditions. With two preparations of the protein that had a high content of alpha 1(I) chains disulfide-linked through the cysteine alpha 1-748, all the large structures formed had a distinctive, highly branched morphology that met one of the formal criteria for a fractal. Preparations with a lower content of disulfide-linked alpha 1(I) chains formed fibrils that were 4 times the diameter of control fibrils. The formation of copolymers was also demonstrated by the observation that the presence of mutated collagens decreased the rate of incorporation of normal collagen into fibrils. In addition, the solution-phase concentration at equilibrium of mixtures of mutated and normal collagens was 5-10-fold greater than that of normal collagen.(ABSTRACT TRUNCATED AT 250 WORDS)
I型前胶原是从一名患有致死性成骨不全变异型的先证者的培养成纤维细胞中纯化得到的。该蛋白质是正常前胶原和突变前胶原的混合物,在一条或两条α1(I)前肽链中存在半胱氨酸取代甘氨酸的情况,其中一些或全部通过α1-748位的半胱氨酸形成二硫键连接。然后,通过用前胶原C蛋白酶将前胶原切割成胶原,在一个从头生成胶原纤维的系统中对该前胶原进行了检测[卡德勒等人(1987年)《生物化学杂志》262卷,15696 - 15701页]。发现在各种实验条件下,突变胶原和正常胶原会形成共聚物。对于两种通过α1-748位半胱氨酸形成二硫键连接的α1(I)链含量高的蛋白质制剂,所有形成的大结构都具有独特的、高度分支的形态,符合分形的一个形式标准。二硫键连接的α1(I)链含量较低的制剂形成的纤维直径是对照纤维的4倍。突变胶原的存在会降低正常胶原掺入纤维的速率,这一观察结果也证明了共聚物的形成。此外,突变胶原与正常胶原混合物在平衡时的溶液相浓度比正常胶原高5 - 10倍。(摘要截短至250字)