A structural and kinetic comparison of proto-oncogenic and oncogenic neu holo-receptors expressed in insect cells.

Receptor Pub Date : 1993-01-01
C M LeVea, J N Myers, W C Dougall, X Qian, M I Greene
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Abstract

The proto-oncogenic and oncogenic forms of the rat neu receptors were expressed in the baculovirus system to characterize their structural and enzymatic differences. The epitopes of their extracellular domains, their molecular weights, and kinase activities were similar to rat neu receptors expressed in fibroblasts. The receptors were partially purified using a phospho-agarose column and were analyzed to compare kinetic parameters using ATP as a substrate. The oncogenic form of the receptor showed a significant increase in Vmax (56%) over the proto-oncogenic form. Structural analysis of these proteins using sucrose gradients showed the oncogenic receptors to have a 62.6% increase in aggregated receptors when compared to the proto-oncogenic receptors. These studies are the first to link enzymatic activation and the physical form of the receptor using isolated receptor species.

昆虫细胞中表达的原致癌和致癌新全息受体的结构和动力学比较。
大鼠新受体的原致癌和致癌形式在杆状病毒系统中表达,以表征它们的结构和酶的差异。它们的胞外结构域表位、分子量和激酶活性与成纤维细胞中表达的大鼠新受体相似。使用磷酸琼脂糖柱部分纯化受体,并使用ATP作为底物分析比较动力学参数。受体的致癌形式显示Vmax比原致癌形式显著增加(56%)。利用蔗糖梯度对这些蛋白进行结构分析,发现与原致癌受体相比,致癌受体的聚集受体增加了62.6%。这些研究是第一次使用分离的受体物种将酶激活和受体的物理形式联系起来。
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