Characterization of the tissue-specific proteoglycans synthesized by chondrocytes from nanomelic chick embryos.
Characterization of the tissue-specific proteoglycans synthesized by chondrocytes from nanomelic chick embryos.
复制标题
纳米鸡胚胎软骨细胞合成的组织特异性蛋白多糖的表征。
DOI：
10.1042/bj2010387
复制
发表时间：
1982
期刊：
影响因子：
--
通讯作者：
Goetinck,PF
中科院分区：
文献类型：
--
作者：
McKeown-Longo,PJ;Goetinck,PF
据报道，来自鸟类突变体奈米利亚的软骨可以合成软骨特异性蛋白聚糖PGS(SC)- 1，含量为正常值的1-2% [McKeown & Goetinck (1979) Dev. Biol. 71, 203-215]。用[35S]硫酸盐内源性标记蛋白多糖，用4 m -盐酸胍从软骨中提取，在1400孔控孔玻璃上进行色谱分析。PGS(SC)-I从这些柱的空隙体积中获得。解离蔗糖-密度梯度分析显示，纳米粒PGS(SC)- 1具有比正常更大的多分散性。从控制孔玻璃的1400空隙体积和蔗糖梯度中分离出来的部分被测试了它们结合软骨蛋白多糖单体的特异性抗体的能力。在所有情况下，正常组分的结合率大于90%，而与纳米组分的结合率在20%至65%之间。PGS(SC)- 1在2500孔控孔玻璃上进行色谱分析，70%的正常蛋白聚糖和25%的突变蛋白聚糖作为聚集体被洗脱。突变体PGS(SC)- 1的硫酸软骨素链在500孔控孔玻璃上比正常的略大。此外，来自纳米软骨的PGS(SC)- 1比来自正常软骨的PGS(SC)- 1更容易在体外被蛋白水解。这表明纳米软骨合成少量软骨特异性蛋白多糖是不正常的。
Cartilage from the avian mutant nanomelia has been reported to synthesize cartilage-specific proteoglycans, PGS(SC)-I, at 1-2% of normal values [McKeown & Goetinck (1979) Dev. Biol. 71, 203-215]. Proteoglycans were endogenously labelled with [35S]sulphate and extracted from cartilage in 4 M-guanidine hydrochloride and chromatographed on controlled-pore glass 1400. PGS(SC)-I was obtained from the void volume of these columns. Dissociative sucrose-density-gradient analysis revealed a greater than normal polydispersity in the nanomelic PGS(SC)-I. Fractions from both the controlled-pore glass 1400 void volume and sucrose gradients were tested for their ability to bind specific antibody against cartilage proteoglycan monomer. In all instances, binding of normal fractions was greater than 90%, whereas binding to nanomelic fractions ranged from 20 to 65%. Chromatography of PGS(SC)-I on controlled-pore glass 2500 resulted in 70% of the normal and 25% of the mutant proteoglycans eluting as aggregates. Chondroitin sulphate chains from mutant PGS(SC)-I appeared slightly larger than normal when chromatographed on controlled-pore glass 500. In addition, PGS(SC)-I from nanomelic cartilage is more susceptible to proteolysis in vitro than the PGS(SC)-I from normal cartilage. This evidence suggests that the small amount of cartilage-specific proteoglycan synthesized by nanomelic cartilage is not normal.