FLUOROMETRIC ANALYSIS-ISOZYMIC FORMS OF CREATINE KINASE

拟议的调查试图将相荧光法应用于一种分析 三级和四级蛋白质结构的相互依存关系 肌酸激酶的二聚体同工酶。根本问题 讨论了亚基折叠和亚基折叠之间相互作用的性质。 亚基组织在非变构的整体构象中， 多聚体蛋白质。荧光分析的最新进展提供了 敏感的和搜索的量化标准，用于比较特定的 结构成分，包括位点到位点的距离、蛋白质旋转 利率、不对称亚单位组织和运动自由 活动站点域。亚基折叠和亚基缔合的动力学 将通过连续测量共振能量传递来进行分析 效率。寿命测量和时间分辨各向异性衰减 相位/调制荧光法是基本的分析方法。 综上所述，这些程序构成了一种实验性的方法 评价三、四级结构在经济发展中的贡献 三维构象的形成和维持及对 肌酸激酶与底物和底物类似物的相互作用。而当 用作非变构齐聚物中亚基相互作用的模型 肌酸激酶表现出几个有趣的特征，是一种 重要的酶研究，因为它在神经肌肉中起着中心作用 能量代谢。这项调查具有代表性的长期目标 更好地理解亚基和低聚物结构的相互依存关系 非变构催化、调节、区隔和稳定 对于生物功能来说是必不可少的。

The proposed investigation seeks to apply phase fluorometry to an analysis of the interdependency of the tertiary and quaternary protein structure of the dimeric isozymes of creatine kinase. The fundamental question addressed is the nature of the interactions between subunit folding and subunit organization in the overall conformation of non-allosteric, multimeric proteins. Recent advances in fluorometric analysis provide sensitive and searching quantitative criteria by which to compare specific structural components including site-site distances, protein rotational rates, asymmetric subunit organization and motional freedom within the active site domain. Kinetics of subunit folding and subunit association will be analyzed by continuous measurement of resonance energy transfer efficiency. Lifetime measurements and time resolved anisotropy decay using phase/modulation fluorometry, constitute essential analytical procedures. Taken together, the procedures constitute an experimental approach to evaluating the contribution of tertiary and quaternary structure in the formation and maintenance of three-dimension conformation and response to interaction with substrate and substrate analog by creatine kinase. While used as a model for subunit interactions in non-allosteric oligomeric enzymes, creatine kinase exhibits several interesting features and is an important enzyme to study because of its central role in neuromuscular energy metabolism. The inquiry is representative the long term goal of better understanding the interdependency of subunit and oligomer structure in non-allosteric catalysis, regulation, compartmentation and stabilization essential for biological function.