McLaughlin C R, Tao Q, Abood M E
Department of Pharmacology/Toxicology, Medical College of Virginia/Virginia Commonwealth University, Richmond 23298-0524.
Nucleic Acids Res. 1994 Dec 11;22(24):5477-83. doi: 10.1093/nar/22.24.5477.
A number of cysteine-rich proteins have recently been isolated by homology screening, differential library screens, and association with other proteins. In this report, we describe the isolation of the rat cysteine-rich protein from a rat brain library during a search for clones with homology to the delta-opioid receptor. One of the cDNAs isolated hybridized to a 1.8 kb mRNA abundantly expressed throughout the rat brain as well as in rat liver. In situ hybridization reveals a wide distribution in rat brain; in particular, abundant hybridization was detected in the hippocampus, cerebellum, habenula, reticular thalamic nucleus and interposed nucleus. Nucleotide sequence analysis of a 1403 bp cDNA clone indicated 77% identity with the cDNA for human cysteine-rich protein (hCRP), that translates into a 99% identity at the amino acid level. The predicted amino acid sequence suggests four zinc fingers, two of the C4 class and two of the C2HC class. This structural motif is characteristic of members of the LIM domain protein family. The mRNA is serum-inducible in Balb/c 3T3 cells. Additional study suggests that its expression is not induced by either NGF treatment of PC12h pheochromocytoma cells, or inflammation-induced injury in the spinal cord at up to 60 min after injury. It does appear to be developmentally expressed in rat brain, consistent with a potential role in neuronal development. The rat CRP clone will be useful for studying the function of CRPs in rodent models.
最近,通过同源性筛选、差异文库筛选以及与其他蛋白质的关联,已经分离出了许多富含半胱氨酸的蛋白质。在本报告中,我们描述了在寻找与δ-阿片受体具有同源性的克隆过程中,从大鼠脑文库中分离大鼠富含半胱氨酸的蛋白质的情况。分离出的一个cDNA与一种在大鼠脑中以及大鼠肝脏中大量表达的1.8 kb mRNA杂交。原位杂交显示在大鼠脑中分布广泛;特别是在海马体、小脑、缰核、丘脑网状核和间位核中检测到大量杂交信号。对一个1403 bp cDNA克隆的核苷酸序列分析表明,它与人类富含半胱氨酸的蛋白质(hCRP)的cDNA具有77%的同一性,在氨基酸水平上则具有99%的同一性。预测的氨基酸序列显示有四个锌指结构,其中两个属于C4类,两个属于C2HC类。这种结构基序是LIM结构域蛋白家族成员的特征。该mRNA在Balb/c 3T3细胞中可被血清诱导。进一步的研究表明,无论是用神经生长因子处理PC12h嗜铬细胞瘤细胞,还是在脊髓损伤后长达60分钟的炎症诱导损伤,都不会诱导其表达。它似乎在大鼠脑中呈发育性表达,这与它在神经元发育中的潜在作用一致。大鼠CRP克隆将有助于在啮齿动物模型中研究CRPs的功能。