ASSESSMENT OF PROCOLLAGEN PROCESSING DEFECTS BY FIBROBLASTS CULTURED IN THE PRESENCE OF DEXTRAN SULFATE
ASSESSMENT OF PROCOLLAGEN PROCESSING DEFECTS BY FIBROBLASTS CULTURED IN THE PRESENCE OF DEXTRAN SULFATE
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DOI：
10.1042/bj2670573
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发表时间：
1990-05-01
影响因子：
4.1
通讯作者：
GOLUB, SB
中科院分区：
文献类型：
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作者：
BATEMAN, JF;GOLUB, SB
在0.01% （w/v）葡聚糖硫酸溶液中培养皮肤成纤维细胞，导致前胶原蛋白完全水解为胶原蛋白。加工主要通过pn中间体进行，这表明c -前肽的切割在加工途径的早期发生。加工过的胶原蛋白与细胞层部分有关。这种诱导前胶原加工的方法被评估用于检测遗传性结缔组织疾病的前胶原加工异常。在两例已知pn -前肽切割缺陷的病例中，I型前胶原加工异常被清楚地证明。在一种情况下，切割缺陷是由于n -蛋白酶活性降低（皮肤稀疏），而在另一种情况下（Ehler' s-Danlos综合征VIIA型），切割位点被删除。在成骨不全（II型）的情况下，I型胶原蛋白的电泳迁移缓慢。由于赖氨酸的过度修饰，-链很容易被证明。正如先前报道的那样，该患者的前胶原蛋白加工效率低下[de Wet， Pihlanjaniemi, Myers, Kelly and Prockop (1983) J. Biol]。化学学报，2004,18(2):521 - 528。因此，这种在葡聚糖硫酸盐存在下培养的方法提供了一种检测前胶原蛋白加工缺陷和电泳异常的简单而快速的方法。
The culture of skin fibroblasts in the presence of 0.01% (w/v) dextran sulphate results in complete proteolytic processing of procollagen to collagen. Processing occurs predominantly via a pN-intermediate, suggesting that C-propeptide cleavage occurs early during the processing pathway. The processed collagen is associated with the cell-layer fraction. This method of inducing procollagen processing was evaluated for use in detecting procollagen processing abnormalities in heritable connective-tissue diseases. Abnormal type I procollagen processing was clearly demonstrated in two cases with known defects of pN-propeptide cleavage. In one, the cleavage deficiency was due to diminished N-proteinase activity (dermatosparaxis) and in the other casee (Ehler''s-Danlos syndrome type VIIA) the cleavage site was deleted. In a case of osteogenesis imperfecta (Type II) the slow electrophoretic migration of type I collagen .alpha.-chains due to over-modification of lysine was readily demonstrated. Inefficient procollagen processing was also evident in this patient, as had been previously reported [de Wet, Pihlanjaniemi, Myers, Kelly and Prockop (1983) J. Biol. Chem. 258, 7721-7728]. Thus this method of culture in the presence of dextran sulphate provides a simple and rapid procedure of the detection of procollagen processing defects and electrophoretic abnormalities.