Development and characterization of fluorescently-labeled myosin light chain kinase calmodulin-binding domain peptides.
Development and characterization of fluorescently-labeled myosin light chain kinase calmodulin-binding domain peptides.
复制标题
荧光标记的肌球蛋白轻链激酶钙调蛋白结合域肽的开发和表征。
DOI：
10.1007/bf01076756
复制
发表时间：
1993
影响因子：
4.3
通讯作者：
Blumenthal,DK
中科院分区：
文献类型：
--
作者：
Blumenthal,DK
钙调蛋白依赖性蛋白激酶如肌球蛋白轻链激酶（MLCK）、钙调蛋白激酶II和磷酸化酶激酶含有负责结合钙调蛋白的特异性序列。这些区域被称为钙调素结合结构域，在许多情况下包含在足够短的序列中，可以通过固相技术合成。化学合成靶酶钙调素结合结构域的能力已经允许使用各种生物物理技术来研究钙调素和钙调素结合结构域肽之间的相互作用。本文综述了基于骨骼肌肌球蛋白轻链激酶钙调素结合域序列的多肽的开发和表征。所提供的数据表明使用荧光标记的肽来研究钙调素-肽相互作用的各个方面，包括结合亲和力、化学计量、特异性、肽构象的变化以及肽-钙调素复合物的热稳定性。这些数据表明，肽表现出许多与钙调素靶酶相互作用的显着特征。因此，荧光标记的肽应作为有用的模型，在分子水平上研究钙调素靶酶的相互作用。
Calmodulin-dependent protein kinases such as myosin light chain kinase (MLCK), calmodulin kinase II, and phosphorylase kinase contain specific sequences responsible for binding calmodulin. These regions are known as calmodulin-binding domains and in many cases are contained within sequences that are short enough to be synthesized by solidphase techniques. The ability to chemically-synthesize target enzyme calmodulin-binding domains has permitted the use of a variety of biophysical techniques to study the interactions between calmodulin and calmodulin-binding domain peptides. The work reviewed here describes the development and characterization of peptides based on the sequence, of the calmodulin-binding domain of skeletal muscle myosin light chain kinase which were labeled with the fluorescent reagent, acrylodan. Data are presented demonstrating the use of fluorescently-labeled peptides to study various aspects of calmodulin-peptide interactions including binding affinity, stoichiometry, specificity, changes in peptide conformation, and thermal stability of the peptide-calmodulin complex. These data indicate the peptides exhibit many of the salient features seen with calmodulin-target enzyme interactions. The fluorescently-labeled peptides should thus serve as useful models for studying calmodulin-target enzyme interactions at the molecular level.