Dutta S K, Bhattacharyya N, Parui R, Verma M
Department of Botany, Howard University, Washington, DC 20059.
Biochem Biophys Res Commun. 1990 Nov 30;173(1):231-9. doi: 10.1016/s0006-291x(05)81046-0.
There is evidence that the gene for gamma-gamma enolase (neuron specific enolase, NSE) is regulated during cell differentiation and development, conserved in a variety of organisms and contains mRNA destabilizing sequences. In order to investigate further the mechanisms of these processes and to obtain large quantity of this protein, the NSE gene was isolated from neuroblastoma cells and cloned in E. coli using standard molecular biology techniques. The NSE gene expression was studied and the expressed protein (recombinant NSE) was characterized extensively. The recombinant NSE behaves like parental NSE in antisera specificity, resistance for chaotropic agents like urea, thermal stability at higher temperatures etc. The physical parameters like secondary structure, hydrophilicity, antigenic index and flexibility of the expressed protein were studied. The results of the present investigation collectively form the basis for initial investigations of how the expression of NSE gene is regulated. This is the first report where the recombinant NSE gene has been characterized so extensively.
有证据表明,γ-γ烯醇化酶(神经元特异性烯醇化酶,NSE)基因在细胞分化和发育过程中受到调控,在多种生物体中保守,并且包含mRNA不稳定序列。为了进一步研究这些过程的机制并获得大量这种蛋白质,使用标准分子生物学技术从神经母细胞瘤细胞中分离出NSE基因并克隆到大肠杆菌中。对NSE基因表达进行了研究,并对表达的蛋白质(重组NSE)进行了广泛表征。重组NSE在抗血清特异性、对尿素等离液剂的抗性、较高温度下的热稳定性等方面表现得与亲本NSE相似。研究了表达蛋白质的二级结构、亲水性、抗原指数和柔韧性等物理参数。本研究结果共同构成了初步研究NSE基因表达如何调控的基础。这是首次对重组NSE基因进行如此广泛表征的报告。