Interaction between the C-terminal region of human myelin basic protein and calmodulin: analysis of complex formation and solution structure.
Interaction between the C-terminal region of human myelin basic protein and calmodulin: analysis of complex formation and solution structure.
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DOI：
10.1186/1472-6807-8-10
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发表时间：
2008-02-19
影响因子：
--
通讯作者：
Kursula, Petri
中科院分区：
文献类型：
--
作者：
Majava, Viivi;Petoukhov, Maxim V.;Hayashi, Nobuhiro;Pirilae, Paeivi;Svergun, Dmitri I.;Kursula, Petri
髓鞘是一种多层膜结构，包裹在轴突周围，使脊椎动物神经冲动的跳跃式传导成为可能。髓鞘碱性蛋白是髓鞘特异性蛋白质中最丰富的蛋白质之一，是一种内在无序的蛋白质，已被证明可以结合钙调蛋白。在这项研究中，我们专注于一个19-mer的合成肽从预测的钙调素结合片段附近的C-末端的人髓鞘碱性蛋白。天然人髓鞘碱性蛋白与钙调素的相互作用通过亲和层析证实。在不同温度下用等温滴定量热法（ITC）测试了髓鞘碱性蛋白肽与钙调素的结合，观察到Kd在低μM范围内，与先前对全长髓鞘碱性蛋白观察到的一样。表面等离子体共振表明，肽结合钙调素，并结合伴随着构象的变化，此外，凝胶过滤色谱法表明在钙调素的存在下的肽的流体动力学半径的减少。NMR光谱用于映射结合区域，主要位于钙调蛋白C-末端叶的疏水口袋内。通过小角X-射线散射获得的溶液结构表明髓鞘碱性蛋白肽结合到钙调蛋白的间隙沟中，而钙调蛋白保持在延伸的构象中。两者合计，我们的研究结果给出了一个详细的结构洞察钙调素与一个主要的髓鞘蛋白，髓鞘碱性蛋白的C-末端片段的相互作用。所使用的19-mer肽主要与钙调素的C-末端叶相互作用，并且伴随结合的构象变化，表明钙调素-靶蛋白相互作用的新模式。钙调素不会塌陷并紧紧地包裹在肽周围;相反，它在溶液结构中保持延伸构象。观察到的亲和力可能是生理相关的，因为神经系统中两种结合配偶体的丰度很高。
The myelin sheath is a multilamellar membrane structure wrapped around the axon, enabling the saltatory conduction of nerve impulses in vertebrates. Myelin basic protein, one of the most abundant myelin-specific proteins, is an intrinsically disordered protein that has been shown to bind calmodulin. In this study, we focus on a 19-mer synthetic peptide from the predicted calmodulin-binding segment near the C-terminus of human myelin basic protein. The interaction of native human myelin basic protein with calmodulin was confirmed by affinity chromatography. The binding of the myelin basic protein peptide to calmodulin was tested with isothermal titration calorimetry (ITC) in different temperatures, and Kd was observed to be in the low μM range, as previously observed for full-length myelin basic protein. Surface plasmon resonance showed that the peptide bound to calmodulin, and binding was accompanied by a conformational change; furthermore, gel filtration chromatography indicated a decrease in the hydrodynamic radius of calmodulin in the presence of the peptide. NMR spectroscopy was used to map the binding area to reside mainly within the hydrophobic pocket of the C-terminal lobe of calmodulin. The solution structure obtained by small-angle X-ray scattering indicates binding of the myelin basic protein peptide into the interlobal groove of calmodulin, while calmodulin remains in an extended conformation. Taken together, our results give a detailed structural insight into the interaction of calmodulin with a C-terminal segment of a major myelin protein, the myelin basic protein. The used 19-mer peptide interacts mainly with the C-terminal lobe of calmodulin, and a conformational change accompanies binding, suggesting a novel mode of calmodulin-target protein interaction. Calmodulin does not collapse and wrap around the peptide tightly; instead, it remains in an extended conformation in the solution structure. The observed affinity can be physiologically relevant, given the high abundance of both binding partners in the nervous system.
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影响因子：
13.8
作者：
Combet, C;Blanchet, C;Deléage, G
通讯作者：
Deléage, G
影响因子：
7.4
作者：
Gestwicki, JE;Hsieh, HV;Pitner, JB
通讯作者：
Pitner, JB
影响因子：
11.2
作者：
Capello, E;Voskuhl, RR;Raine, CS
通讯作者：
Raine, CS
影响因子：
4.2
作者：
Galiano, M. R.;Andrieux, A.;Hallak, M. E.
通讯作者：
Hallak, M. E.
DOI：
10.1046/j.1432-1327.1998.2550060.x
发表时间：
1998-07-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Göhring, W;Sasaki, T;Timpl, R
通讯作者：
Timpl, R