Fisher L W, Termine J D, Young M F
Bone Research Branch, National Institute of Dental Research, Bethesda, Maryland 20892.
J Biol Chem. 1989 Mar 15;264(8):4571-6.
The small proteoglycans (PG) of bone consist of two different molecular species: one containing one chondroitin sulfate chain (PG II) and the other, two chains (PG I). These two proteoglycans are found in many connective tissues and have Mr = 45,000 core proteins with clear differences in their NH2-terminal sequences. Using antisera produced against synthetic peptides derived from the human PG I and PG II NH2 termini, we have isolated several cDNA clones from a lambda gt11 expression library made against mRNA isolated from human bone-derived cells. The clones, which reacted with antisera to the PG II peptide, were sequenced and found to be identical with the PG II class of proteoglycan from human fibroblasts known as PG-40 or decorin. The clones reacting to the PG I antisera, however, had a unique sequence. The derived protein sequence of PG I showed sufficient homology with the PG II sequence (55% of the amino acids are identical, with most others involving chemically similar amino acid substitutions) to strongly suggest that the two proteins were the result of a gene duplication. PG II (decorin) contains one attached glycosaminoglycan chain, while PG I probably contains two chains. For this reason, we suggest that PG I be called biglycan. The biglycan protein sequence contains 368 residues (Mr = 42,510 for the complete sequence and Mr = 37,983 for the secreted form) that appears to consist predominantly of a series of 12 tandem repeats of 24 residues. The repeats are recognized by their conserved leucines (and leucine-like amino acids) in positions previously reported for a diverse collection of proteins (none of which is thought to be proteoglycans) including: two morphogenic proteins (toll and chaoptin) in the fruit fly; a yeast adenylate cyclase; and two human proteins, the von Willebrand Factor-binding platelet membrane protein, GPIb, and a rare serum protein, leucine-rich glycoprotein.
骨组织中的小蛋白聚糖(PG)由两种不同的分子类型组成:一种含有一条硫酸软骨素链(PG II),另一种含有两条链(PG I)。这两种蛋白聚糖存在于许多结缔组织中,其核心蛋白的分子量为45,000,在氨基末端序列上有明显差异。我们利用针对源自人PG I和PG II氨基末端的合成肽产生的抗血清,从一个λgt11表达文库中分离出了几个cDNA克隆,该文库是根据从人骨来源细胞中分离的mRNA构建的。与PG II肽抗血清发生反应的克隆经测序后,发现与来自人成纤维细胞的PG II类蛋白聚糖(称为PG - 40或核心蛋白聚糖)相同。然而,与PG I抗血清发生反应的克隆具有独特的序列。PG I的推导蛋白序列与PG II序列具有足够的同源性(55%的氨基酸相同,大多数其他氨基酸涉及化学性质相似的氨基酸替换),这强烈表明这两种蛋白质是基因复制的结果。PG II(核心蛋白聚糖)含有一条连接的糖胺聚糖链,而PG I可能含有两条链。因此,我们建议将PG I称为双糖链蛋白聚糖。双糖链蛋白聚糖的蛋白序列包含368个残基(完整序列的分子量为42,510,分泌形式的分子量为37,983),似乎主要由一系列12个24残基的串联重复序列组成。这些重复序列通过其保守的亮氨酸(和类亮氨酸氨基酸)在先前报道的多种蛋白质(其中没有一种被认为是蛋白聚糖)中的位置得以识别,这些蛋白质包括:果蝇中的两种形态发生蛋白(Toll和chaoptin);一种酵母腺苷酸环化酶;以及两种人类蛋白质,即与血管性血友病因子结合的血小板膜蛋白GPIb和一种罕见的血清蛋白,富含亮氨酸的糖蛋白。