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Title: Identification and localization of bound internal water in the solution structure of interleukin 1. beta. by heteronuclear three-dimensional sup 1 H rotating-frame Overhauser sup 15 N- sup 1 H multiple quantum coherence NMR spectroscopy

Abstract

The presence and location of bound internal water molecules in the solution structure of interleukin 1{beta} have been investigated by means of three-dimensional {sup 1}H rotating-frame Overhauser {sup 1}H-{sup 15}N multiple quantum coherence spectroscopy (ROESY-HMQC). In this experiment through-space rotating-frame Overhauser (ROE) interactions between NH protons and bound water separated by {le}3.5{angstrom} are clearly distinguished from chemical exchange effects, as the cross-peaks for these two processes are of opposite sign. The identification of ROEs between NH protons and water is rendered simple by spreading out the spectrum into a third dimensional according to the {sup 15}N chemical shift of the directly bonded nitrogen atoms. By this means, the problems that prevent, in all but a very few limited cases, the interpretation, identification, and assignment of ROE peaks between NH protons and water in a 2D {sup 1}H-{sup 1}H ROESY spectrum of a large protein such as interleukin 1{beta}, namely, extensive NH chemical shift degeneracy and ROE peaks obscured by much stronger chemical exchange peaks, are completely circumvented. We demonstrate the existence of 15 NH protons that are close to bound water molecules. From an examination of the crystal structure of interleukin, the results can be attributed to 11 water moleculesmore » that are involved in interactions bridging hydrogen-bonding interactions with backbone amide and carbonyl groups which stabilize the 3-fold pseudosymmetric topology of interleukin 1{beta} and thus constitute an integral part of the protein structure in solution.« less

Authors:
; ; ;  [1]
  1. National Institutes of Health, Bethesda, MD (USA)
Publication Date:
OSTI Identifier:
6038729
Resource Type:
Journal Article
Journal Name:
Biochemistry; (USA)
Additional Journal Information:
Journal Volume: 29:24; Journal ID: ISSN 0006-2960
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; LYMPHOKINES; NUCLEAR MAGNETIC RESONANCE; WATER; AQUEOUS SOLUTIONS; CHEMICAL SHIFT; FOURIER TRANSFORMATION; LABELLED COMPOUNDS; NITROGEN 15; OVERHAUSER EFFECT; PROTONS; BARYONS; DISPERSIONS; ELEMENTARY PARTICLES; FERMIONS; GROWTH FACTORS; HADRONS; HYDROGEN COMPOUNDS; INTEGRAL TRANSFORMATIONS; ISOTOPES; LIGHT NUCLEI; MAGNETIC RESONANCE; MITOGENS; MIXTURES; NITROGEN ISOTOPES; NUCLEI; NUCLEONS; ODD-EVEN NUCLEI; ORGANIC COMPOUNDS; OXYGEN COMPOUNDS; PROTEINS; RESONANCE; SOLUTIONS; STABLE ISOTOPES; TRANSFORMATIONS; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Clore, G M, Bax, A, Wingfield, P T, and Gronenborn, A M. Identification and localization of bound internal water in the solution structure of interleukin 1. beta. by heteronuclear three-dimensional sup 1 H rotating-frame Overhauser sup 15 N- sup 1 H multiple quantum coherence NMR spectroscopy. United States: N. p., 1990. Web. doi:10.1021/bi00476a004.
Clore, G M, Bax, A, Wingfield, P T, & Gronenborn, A M. Identification and localization of bound internal water in the solution structure of interleukin 1. beta. by heteronuclear three-dimensional sup 1 H rotating-frame Overhauser sup 15 N- sup 1 H multiple quantum coherence NMR spectroscopy. United States. https://doi.org/10.1021/bi00476a004
Clore, G M, Bax, A, Wingfield, P T, and Gronenborn, A M. 1990. "Identification and localization of bound internal water in the solution structure of interleukin 1. beta. by heteronuclear three-dimensional sup 1 H rotating-frame Overhauser sup 15 N- sup 1 H multiple quantum coherence NMR spectroscopy". United States. https://doi.org/10.1021/bi00476a004.
@article{osti_6038729,
title = {Identification and localization of bound internal water in the solution structure of interleukin 1. beta. by heteronuclear three-dimensional sup 1 H rotating-frame Overhauser sup 15 N- sup 1 H multiple quantum coherence NMR spectroscopy},
author = {Clore, G M and Bax, A and Wingfield, P T and Gronenborn, A M},
abstractNote = {The presence and location of bound internal water molecules in the solution structure of interleukin 1{beta} have been investigated by means of three-dimensional {sup 1}H rotating-frame Overhauser {sup 1}H-{sup 15}N multiple quantum coherence spectroscopy (ROESY-HMQC). In this experiment through-space rotating-frame Overhauser (ROE) interactions between NH protons and bound water separated by {le}3.5{angstrom} are clearly distinguished from chemical exchange effects, as the cross-peaks for these two processes are of opposite sign. The identification of ROEs between NH protons and water is rendered simple by spreading out the spectrum into a third dimensional according to the {sup 15}N chemical shift of the directly bonded nitrogen atoms. By this means, the problems that prevent, in all but a very few limited cases, the interpretation, identification, and assignment of ROE peaks between NH protons and water in a 2D {sup 1}H-{sup 1}H ROESY spectrum of a large protein such as interleukin 1{beta}, namely, extensive NH chemical shift degeneracy and ROE peaks obscured by much stronger chemical exchange peaks, are completely circumvented. We demonstrate the existence of 15 NH protons that are close to bound water molecules. From an examination of the crystal structure of interleukin, the results can be attributed to 11 water molecules that are involved in interactions bridging hydrogen-bonding interactions with backbone amide and carbonyl groups which stabilize the 3-fold pseudosymmetric topology of interleukin 1{beta} and thus constitute an integral part of the protein structure in solution.},
doi = {10.1021/bi00476a004},
url = {https://www.osti.gov/biblio/6038729}, journal = {Biochemistry; (USA)},
issn = {0006-2960},
number = ,
volume = 29:24,
place = {United States},
year = {Tue Jun 19 00:00:00 EDT 1990},
month = {Tue Jun 19 00:00:00 EDT 1990}
}