Characterization of a cDNA for chicken osteopontin: expression during bone development, osteoblast differentiation, and tissue distribution.
Characterization of a cDNA for chicken osteopontin: expression during bone development, osteoblast differentiation, and tissue distribution.
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DOI：
10.1021/bi00223a029
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发表时间：
1991-03
期刊：
影响因子：
2.9
通讯作者：
M. Moore;Y. Gotoh;K. Rafidi;L. Gerstenfeld
中科院分区：
文献类型：
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作者：
M. Moore;Y. Gotoh;K. Rafidi;L. Gerstenfeld
鸡骨磷蛋白（约66-kDa BPP）是骨的主要非胶原组分，并且是由培养的鸡胚成骨细胞合成的主要磷蛋白[Gotoh，Y.，格斯滕费尔德湖C.的方法，& Glimcher，M. J.（1990）Eur. 87，49-58]。从鸡胚骨mRNA表达文库中分离出该蛋白的cDNA克隆。从cDNA序列推导出264个氨基酸的完整的一级蛋白质序列，包括16个氨基酸的信号肽序列，并以4个读框内终止序列终止。序列比对表明，约35%的总体相似性，在蛋白质序列之间的鸟类约66 kDa的BPP和哺乳动物蛋白质骨桥蛋白，而在核苷酸水平上观察到60%的相似性。这一序列的特点，表现出最大的相似性，哺乳动物骨桥蛋白包括一个区域，其中7个连续的9个残基是天冬氨酸，整合素介导的细胞结合的识别序列（-精氨酸-甘氨酸-天冬氨酸），和四个可能的识别序列的磷酸化酪蛋白激酶II。杂交分析表明，主要在骨和肾中发现的1.5 kb的信息。mRNA在佛波酯处理的软骨细胞的原代培养物中是可诱导的，其在正常生长条件下不显示表达。在体内和体外成骨细胞分化过程中观察到时间诱导，从而表明在成骨细胞发育过程中约66-kDa BPP的调节受转录控制。总之，蛋白质的一级结构和其生物学特性表明，它是哺乳动物蛋白骨桥蛋白的鸟类同源物。
The chicken bone phosphoprotein (approximately 66-kDa BPP) is a major noncollagenous component of bone and is the major phosphoprotein synthesized by cultured chicken embryo osteoblasts [Gotoh, Y., Gerstenfeld, L. C., & Glimcher, M. J. (1990) Eur. J. Biochem. 87, 49-58]. A cDNA clone for this protein was isolated from an expression library made from embryonic chicken bone mRNA. The complete primary protein sequence of 264 amino acids was deduced from the cDNA sequence inclusive of a 16 amino acid signal peptide sequence and terminated by 4 in-frame stop sequences. A sequence alignment indicated an approximate 35% overall similarity in protein sequence between the avian approximately 66-kDa BPP and the mammalian protein osteopontin, while at the nucleotide level 60% similarity was observed. Features of this sequence which showed the greatest similarity to mammalian osteopontin included a region in which seven of nine consecutive residues are aspartic acid, a recognition sequence for integrin-mediated cell binding (-Arg-Gly-Asp), and four possible recognition sequences for phosphorylation by casein kinase II. Hybridization analysis indicated a message of 1.5 kb found predominantly in bone and kidney. The mRNA was inducible in phorbol ester treated primary cultures of chondrocytes which show no expression under normal growth conditions. A temporal induction was seen during osteoblastic differentiation both in vivo and in vitro, thus suggesting that regulation of the approximately 66-kDa BPP is under transcriptional control during osteoblast development. In summary, both the protein's primary structure and its biological features suggest that it is the avian homologue to mammalian protein osteopontin.