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

Journal Article · Tue Jun 19 00:00:00 EDT 1990 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00476a004· OSTI ID:6038729

Clore, G M; Bax, A; Wingfield, P T; Gronenborn, A M ^[1]

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1. National Institutes of Health, Bethesda, MD (USA)

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.

• OSTI ID: 6038729
• Journal Information: Biochemistry; (USA), Vol. 29:24; ISSN 0006-2960
• Country of Publication: United States
• Language: English

Related Subjects

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