ASSIGNMENT OF H-1, N-15, AND C-13 RESONANCES, IDENTIFICATION OF ELEMENTS OF SECONDARY STRUCTURE AND DETERMINATION OF THE GLOBAL FOLD OF THE DNA-BINDING DOMAIN OF GAL4

被引:31
作者
SHIRAKAWA, M
FAIRBROTHER, WJ
SERIKAWA, Y
OHKUBO, T
KYOGOKU, Y
WRIGHT, PE
机构
[1] OSAKA UNIV, INST PROT RES, SUITA, OSAKA 565, JAPAN
[2] SCRIPPS RES INST, DEPT MOLEC BIOL, LA JOLLA, CA 92037 USA
[3] PROT ENGN RES INST, SUITA, OSAKA 565, JAPAN
关键词
D O I
10.1021/bi00060a004
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Almost complete assignments of the H-1, N-15, and aliphatic C-13 resonances of the 62-residue N-terminal DNA-binding domain of GAL4 [GAL4(62)] have been obtained using a combination of two-dimensional homonuclear and two- and three-dimensional double- and triple-resonance heteronuclear NMR methods. The sequential NOE connectivities, amide proton exchange measurements, and C-13alpha chemical shift data indicate the presence of two short alpha-helices in the N-terminal half of the polypeptide. Residues 1-9 and 41-62 appear to be unstructured and flexible in solution. Analysis of the C-13alpha chemical shifts also revealed a significant downfield shift of approximately +3 ppm, relative to random-coil values, for the four nonbridging Zn(II) ligands, Cys 14, 21, 31, and 38. Interestingly, no such correlation was observed for the two bridging ligands, Cys 11 and 28. Preliminary structure calculations using a subset of distance restraints derived from three-dimensional H-1-N-15 and H-1-C-13 NOESY-HSQC spectra are consistent with the recently reported solution structures of Zn(II)2GAL4(7-49) [Kraulis, P., et al. (1992) Nature 356, 448-450] and of Cd(II)2GAL4(65) [Baleja, J. D., et al. (1992) Nature 356, 450-453].
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页码:2144 / 2153
页数:10
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