Refined solution structure and backbone dynamics of 15N-labeled C12A-p8MTCP studied by NMR relaxation
Refined solution structure and backbone dynamics of 15N-labeled C12A-p8MTCP studied by NMR relaxation
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通过 NMR 弛豫研究 15N 标记的 C12A-p8MTCP 的精细溶液结构和主链动力学
DOI：
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发表时间：
1999
影响因子：
2.7
通讯作者：
C. Roumestand
中科院分区：
文献类型：
--
作者：
P. Barthe;L. Chiche;N. Declerck;M. Delsuc;J. Lefèvre;T. Malliavin;J. Mispelter;M. Stern;J. Lhoste;C. Roumestand
MTCP 1（用于成熟T细胞增殖）是在具有成熟表型的罕见白血病组中明确鉴定的第一个基因。由MTCP 1癌基因编码的人p8 MTCP蛋白的三维溶液结构先前已通过600 MHz的同源质子二维NMR方法确定：它由包含三个α-螺旋的原始支架组成，与新的半胱氨酸基序相关。其中两个螺旋通过两个二硫键共价配对，形成类似于反平行卷曲螺旋的α-发夹。第三个螺旋的方向大致平行于α-反平行基序所定义的平面，并且看起来不太明确。为了更深入地了解这种新支架的细节，我们用氮-15均匀地标记了这种蛋白质的突变体（C12 A-p8 MTCP 1），其中位置12处的未结合的半胱氨酸已被丙氨酸残基取代，从而允许重组蛋白的可再现的高产率。精确的结构得益于从15 N编辑的3D实验中提取的211个额外的NOE，以及一组几乎完整的φ角约束，允许估计蛋白质结构部分的螺旋含量。此外，15 N自旋弛豫时间和异核15 N1 HNOE的测量为蛋白质骨架的动力学提供了额外的见解。利用J（0）与J（ω）之间的线性相关分析来解释弛豫参数。看来，在螺旋III中看到的明显的相对无序并不仅仅是由于缺乏实验限制，而是与这一段中亚纳秒运动的实质性贡献有关。
MTCP1 (for Mature-T-Cell Proliferation) was the first gene unequivocally identified in the group of uncommon leukemias with a mature phenotype. The three-dimensional solution structure of the human p8MTCP protein encoded by the MTCP1 oncogene has been previously determined by homonuclear proton two-dimensional NMR methods at 600 MHz: it consists of an original scaffold comprising three α-helices, associated with a new cysteine motif. Two of the helices are covalently paired by two disulfide bridges, forming an α-hairpin which resembles an antiparallel coiled-coil. The third helix is orientated roughly parallel to the plane defined by the α-antiparallel motif and appears less well defined. In order to gain more insight into the details of this new scaffold, we uniformly labeled with nitrogen-15 a mutant of this protein (C12A-p8MTCP1) in which the unbound cysteine at position 12 has been replaced by an alanine residue, thus allowing reproducibly high yields of recombinant protein. The refined structure benefits from 211 additional NOEs, extracted from 15N-edited 3D experiments, and from a nearly complete set of φ angular restraints allowing the estimation of the helical content of the structured part of the protein. Moreover, measurements of 15 N spin relaxation times and heteronuclear 15 N1HNOEs provided additional insights into the dynamics of the protein backbone. The analysis of the linear correlation between J(0) and J(ω) was used to interpret relaxation parameters. It appears that the apparent relative disorder seen in helix III is not simply due to a lack of experimental constraints, but associated with substantial contributions of sub-nanosecond motions in this segment.
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影响因子：
2.9
作者：
Lefevre, JF;Dayie, KT;Wagner, G
通讯作者：
Wagner, G
影响因子：
2.9
作者：
PENG, JW;WAGNER, G
通讯作者：
WAGNER, G
影响因子：
5.6
作者：
BRADLEY, EK;THOMASON, JF;KUNTZ, ID
通讯作者：
KUNTZ, ID
通过异核弛豫测量和降低的谱密度映射探测 villin 14T 内的局部迁移率。
DOI：
10.1021/bi951933o
发表时间：
1996
期刊：
Biochemistry.
影响因子：
--
作者：
Markus,MA;Dayie,KT;Matsudaira,P;Wagner,G
通讯作者：
Wagner,G