Solution structure of a naturally-occurring zinc-peptide complex demonstrates that the N-terminal zinc-binding module of the Lasp-1 LIM domain is an independent folding unit
Solution structure of a naturally-occurring zinc-peptide complex demonstrates that the N-terminal zinc-binding module of the Lasp-1 LIM domain is an independent folding unit
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DOI：
10.1021/bi961149j
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发表时间：
1996-10-01
期刊：
影响因子：
2.9
通讯作者：
Otting, G
中科院分区：
文献类型：
--
作者：
Hammarstrom, A;Berndt, KD;Otting, G
合成的肽ZF-1和锌之间的1：1的复合物的三维溶液结构由H-1 NMR光谱确定。最初从猪肠中分离的肽在序列上与属于LIM结构域蛋白家族的人蛋白Lasp-1的30个N-末端氨基酸残基相同。最后一组20个能量精炼的NMR构象异构体相对于残基3-30的骨架原子的平均结构具有0.55埃的平均rmsd。没有锌原子限制的计算明确地将Cys 5、Cys 8、His 26和Cys 29鉴定为锌配位残基。LIM结构域由两个连续的锌结合模块组成，并且ZF-1（-）锌络合物的NMR结构是孤立模块结构的第一个实例。与鸡CRP和大鼠GRIP的第二个LIM结构域的N-末端锌结合模块的已知结构的比较，其中ZF-1分别具有50%和43%的序列同一性，支持LIM结构域的锌结合模块具有保守的结构基序并识别结构多样性的局部区域的概念。相似之处包括保守的锌配位残基，涉及Cys 5和Cys 8的rubredoxin关节，以及与其他锌指结构域观察到的更常见的N-末端配位相比，组氨酸N-δ与锌离子的配位。ZF-1（-）锌复合物的本结构测定建立了LIM结构域的N-末端一半作为独立的折叠单元。LIM结构域的N-和C-末端锌结合模块的结构相似性，尽管有限的序列同一性，导致在LIM结构域中的单个锌结合基序的建议。坐标可从Brookhaven蛋白质数据库条目1 ZFO获得。
The three-dimensional solution structure of the 1:1 complex between the synthetic peptide ZF-1 and zinc was determined by H-1 NMR spectroscopy. The peptide, initially isolated from pig intestines, is identical in sequence to the 30 N-terminal amino acid residues of the human protein Lasp-1 belonging to the LIM domain protein family. The final set of 20 energy-refined NMR conformers has an average rmsd relative to the mean structure of 0.55 Angstrom for the backbone atoms of residues 3-30, Calculations without zinc atom constraints unambiguously identified Cys 5, Cys 8, His 26, and Cys 29 as the zinc-coordinating residues. LIM domains consist of two sequential zinc-binding modules and the NMR structure of the ZF-1(-)zinc complex is the first example of a structure of an isolated module. Comparison with the known structures of the N-terminal zinc-binding modules of both the second LIM domain of chicken CRP and rat GRIP with which ZF-1 shares 50% and 43% sequence identity, respectively, supports the notion that the zinc-binding modules of the LIM domain have a conserved structural motif and identifies local regions of structural diversity. The similarities include conserved zinc-coordinating residues, a rubredoxin knuckle involving Cys 5 and Cys 8, and the coordination of the zinc ion by histidine N-delta in contrast to the more usual coordination by N-epsilon observed for other zinc-finger domains, The present structure determination of the ZF-1(-)zinc complex establishes the N-terminal half of a LIM domain as an independent folding unit. The structural similarities of N- and C-terminal zinc-binding modules of the LIM domains, despite limited sequence identity, lead to the proposal of a single zinc-binding motif in LIM domains. The coordinates are available from the Brookhaven protein data bank, entry 1ZFO.