Application of HN(C)N to rapid estimation of 1J(N-Cα) coupling constants correlated to ψ torsion angles in proteins:: implication to structural genomics

被引:6
作者
Bhavesh, NS
Chatterjee, A
Panchal, SC
Hosur, RV
机构
[1] Tata Inst Fundamental Res, Dept Chem Sci, Bombay 400005, Maharashtra, India
[2] UT SW Med Ctr, Dept Biochem, Dallas, TX 75390 USA
关键词
HN(C)N; 1J(N-C alpha) coupling; torsion angles; FK506 binding protein; structural genomics;
D O I
10.1016/j.bbrc.2003.09.237
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
We recently described a triple resonance experiment, HN(C)N, for sequential correlation of H-N and N-15 atoms in (N-15, C-13) labeled proteins [J. Biomol. NMR. 20 (2001) 135]. Here, we describe an approach based on this experiment for estimation of one bond N-C-alpha J-couplings in medium size labeled proteins, which seem to show good correlations with psi torsion angles along the protein backbone. The approach uses the ratio of the intensities of the sequential and diagonal peaks in the F-2-F-3 planes of the HN(C)N spectrum. The reliability of the approach has been demonstrated using a short peptide wherein the coupling constants have been measured by the present method and also independently from peak splittings in HSQC spectra. The two results agree within 10%. The applicability of the procedure to proteins has been demonstrated using doubly labeled FK506 binding protein (FKBP, molecular mass similar to12 kDa). Coupling constant estimates have been obtained for 62 out of 100 non-proline residues and they show a correlation with psi torsion angles, as has been reported before. This semi-quantitative application of HN(C)N extends the significance of the experiment especially, in the context of structural genomics, since the single experiment, not only provides a great enhancement in the speed of resonance assignment, but also provides quantitative structural information. (C) 2003 Elsevier Inc. All rights reserved.
引用
收藏
页码:678 / 684
页数:7
相关论文
共 23 条
[1]  
BAX A, 1994, METHOD ENZYMOL, V239, P79
[2]   The Protein Data Bank [J].
Berman, HM ;
Westbrook, J ;
Feng, Z ;
Gilliland, G ;
Bhat, TN ;
Weissig, H ;
Shindyalov, IN ;
Bourne, PE .
NUCLEIC ACIDS RESEARCH, 2000, 28 (01) :235-242
[3]   NMR identification of local structural preferences in HIV-1 protease tethered heterodimer in 6 M guanidine hydrochloride [J].
Bhavesh, NS ;
Panchal, SC ;
Mittal, R ;
Hosur, RV .
FEBS LETTERS, 2001, 509 (02) :218-224
[4]   An efficient high-throughput resonance assignment procedure for structural genomics and protein folding research by NMR [J].
Bhavesh, NS ;
Panchal, SC ;
Hosur, RV .
BIOCHEMISTRY, 2001, 40 (49) :14727-14735
[5]   PRECISE VICINAL COUPLING-CONSTANTS 3JHN-ALPHA IN PROTEINS FROM NONLINEAR FITS OF J-MODULATED [N-15,H-1]-COSY EXPERIMENTS [J].
BILLETER, M ;
NERI, D ;
OTTING, G ;
QIAN, YQ ;
WUTHRICH, K .
JOURNAL OF BIOMOLECULAR NMR, 1992, 2 (03) :257-274
[6]   A novel protocol based on HN(C)N for rapid resonance assignment in (15N,13C) labeled proteins:: implications to structural genomics [J].
Chatterjee, A ;
Bhavesh, NS ;
Panchal, SC ;
Hosur, RV .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2002, 293 (01) :427-432
[7]   N-15 NMR RELAXATION STUDIES OF THE FK506 BINDING-PROTEIN - BACKBONE DYNAMICS OF THE UNCOMPLEXED RECEPTOR [J].
CHENG, JW ;
LEPRE, CA ;
CHAMBERS, SP ;
FULGHUM, JR ;
THOMSON, JA ;
MOORE, JM .
BIOCHEMISTRY, 1993, 32 (35) :9000-9010
[8]  
DELAGLIO F, 1991, Journal of Biomolecular NMR, V1, P439, DOI 10.1007/BF02192865
[9]   CALCULATIONS OF ONE-BOND, 2-BOND AND 3-BOND NUCLEAR SPIN-SPIN COUPLINGS IN A MODEL PEPTIDE AND CORRELATIONS WITH EXPERIMENTAL-DATA [J].
EDISON, AS ;
MARKLEY, JL ;
WEINHOLD, F .
JOURNAL OF BIOMOLECULAR NMR, 1994, 4 (04) :519-542
[10]   ESTIMATES OF PHI-TORSION AND PSI-TORSION ANGLES IN PROTEINS FROM ONE-BOND, 2-BOND AND 3-BOND NUCLEAR SPIN-SPIN COUPLINGS - APPLICATION TO STAPHYLOCOCCAL NUCLEASE [J].
EDISON, AS ;
WEINHOLD, F ;
WESTLER, WM ;
MARKLEY, JL .
JOURNAL OF BIOMOLECULAR NMR, 1994, 4 (04) :543-551