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Automated protein fold determination using a minimal NMR constraint strategy
Author(s) -
Zheng Deyou,
Huang Yuanpeng J.,
Moseley Hunter N.B.,
Xiao Rong,
Aramini James,
Swapna G.V.T.,
Montelione Gaetano T.
Publication year - 2003
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.0300203
Subject(s) - chemistry , two dimensional nuclear magnetic resonance spectroscopy , nuclear magnetic resonance spectroscopy , side chain , nmr spectra database , protonation , carbon 13 nmr satellite , nuclear overhauser effect , nuclear magnetic resonance , biological system , spectral line , stereochemistry , fluorine 19 nmr , physics , polymer , ion , organic chemistry , astronomy , biology
Determination of precise and accurate protein structures by NMR generally requires weeks or even months to acquire and interpret all the necessary NMR data. However, even medium‐accuracy fold information can often provide key clues about protein evolution and biochemical function(s). In this article we describe a largely automatic strategy for rapid determination of medium‐accuracy protein backbone structures. Our strategy derives from ideas originally introduced by other groups for determining medium‐accuracy NMR structures of large proteins using deuterated, 13 C‐, 15 N‐enriched protein samples with selective protonation of side‐chain methyl groups ( 13 CH 3 ). Data collection includes acquiring NMR spectra for automatically determining assignments of backbone and side‐chain 15 N, H N resonances, and side‐chain 13 CH 3 methyl resonances. These assignments are determined automatically by the program AutoAssign using backbone triple resonance NMR data, together with Spin System Type Assignment Constraints (STACs) derived from side‐chain triple‐resonance experiments. The program AutoStructure then derives conformational constraints using these chemical shifts, amide 1 H/ 2 H exchange, nuclear Overhauser effect spectroscopy (NOESY), and residual dipolar coupling data. The total time required for collecting such NMR data can potentially be as short as a few days. Here we demonstrate an integrated set of NMR software which can process these NMR spectra, carry out resonance assignments, interpret NOESY data, and generate medium‐accuracy structures within a few days. The feasibility of this combined data collection and analysis strategy starting from raw NMR time domain data was illustrated by automatic analysis of a medium accuracy structure of the Z domain of Staphylococcal protein A.