NMR STRUCTURE DETERMINATION OF THE ESCHERICHIA-COLI DNAJ MOLECULAR CHAPERONE - SECONDARY STRUCTURE AND BACKBONE FOLD OF THE N-TERMINAL REGION (RESIDUES-2-108) CONTAINING THE HIGHLY CONSERVED J-DOMAIN
NMR STRUCTURE DETERMINATION OF THE ESCHERICHIA-COLI DNAJ MOLECULAR CHAPERONE - SECONDARY STRUCTURE AND BACKBONE FOLD OF THE N-TERMINAL REGION (RESIDUES-2-108) CONTAINING THE HIGHLY CONSERVED J-DOMAIN
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DOI：
10.1073/pnas.91.24.11343
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发表时间：
1994-11-22
影响因子：
11.1
通讯作者：
WUTHRICH, K
中科院分区：
文献类型：
--
作者：
SZYPERSKI, T;PELLECCHIA, M;WUTHRICH, K
来自大肠杆菌的DnaJ是一种由376个氨基酸组成的蛋白质，它与DnaK和GrpE一起作为伴侣机器起作用。残基2-108的n端片段DnaJ-(2-108)保留了全长蛋白的许多活性，并包含一个结构蛾，残基2-72的J结构域，在蛋白质超家族中高度保守。本文采用核磁共振波谱法测定了DnaJ（2-108）的二级结构和三维多肽主链折叠。通过使用C-13/N-15双标记DnaJ-(2-108)，获得了H-1、N-15、C-13（α）和C-13（β）的几乎完整的序列特异性定位，并确定了约40%的外周脂肪碳共振。J结构域6-11、18-31、41-55和61-68残基多肽段的4个cu螺旋通过序列和中程核Overhauser效应被鉴定出来。对于J畴，用DIANA程序计算了536个核Overhauser效应上距离约束和52个自旋-自旋耦合常数的三维结构。多肽主链折叠的特点是形成一个反平行的两个长螺旋束，残基18-31和41-55，由同源J结构域序列高度保守的疏水侧链核心稳定。残基77 ~ 108之间的Gly/ ph富区在溶液中是灵活无序的。
DnaJ from Escherichia coli is a 376-amino acid protein that functions in conjunction with DnaK and GrpE as a chaperone machine. The N-terminal fragment of residues 2-108, DnaJ-(2-108), retains many of the activities of the full-length protein and contains a structural moth, the J domain Of residues 2-72, which is highly conserved in a superfamily of proteins. In this paper, NMR spectroscopy was used to determine the secondary structure and the three-dimensional polypeptide backbone fold of DnaJ (2-108). By using C-13/N-15 doubly labeled DnaJ-(2-108), nearly complete sequence-specific assignments were obtained for H-1, N-15, C-13(alpha), and C-13(beta), and about 40% of the peripheral aliphatic carbon resonances were also assigned. Four cu-helices in polypeptide segments of residues 6-11, 18-31, 41-55, and 61-68 in the J domain were identified by sequential and medium-range nuclear Overhauser effects. For the J domain, the three-dimensional structure was calculated with the program DIANA from an input of 536 nuclear Overhauser effect upper-distance constraints and 52 spin-spin coupling constants. The polypeptide backbone fold is characterized by the formation of an antiparallel bundle of two long helices, residues 18-31 and 41-55, which is stabilized by a hydrophobic core of side chains that are highly conserved In homologous J domain sequences. The Gly/Phe-rich region from residues 77 to 108 is flexibly disordered in solution.