Complete resonance assignment for the polypeptide backbone of interleukin 1. beta. using three-dimensional heteronuclear NMR spectroscopy

Journal Article · Tue Apr 10 00:00:00 EDT 1990 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00466a018· OSTI ID:6170376

Driscoll, P C; Clore, G M; Marion, D; Wingfield, P T ^[1]

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1. Glaxo Institute for Molecular Biology (Switzerland)

The complete sequence-specific assignment of the {sup 15}N and {sup 1}H backbone resonances of the NMR spectrum of recombinant human interleukin 1{beta} has been obtained by using primarily {sup 15}N-{sup 1}H heteronuclear three-dimensional (3D) NMR techniques in combination with {sup 15}N-{sup 1}H heteronuclear and {sup 1}H homonuclear two-dimensional NMR. The fingerprint region of the spectrum was analyzed by using a combination of 3D heteronuclear {sup 1}H Hartmann-Hahn {sup 15}N-{sup 1}H multiple quantum coherence (3D HOHAHA-HMQC) and 3D heteronuclear {sup 1}H nuclear Overhauser {sup 15}N-{sup 1}H multiple quantum coherence (3D NOESY-HMQC) spectroscopies. The authors show that the problems of amide NH and C{sup {alpha}}H chemical shift degeneracy that are prevalent for proteins of the size are readily overcome by using the 3D heteronuclear NMR technique. A doubling of some peaks in the spectrum was found to be due to N-terminal heterogeneity of the {sup 15}N-labeled protein, corresponding to a mixture of wild-type and des-Ala-1-interleukin 1{beta}. The complete list of {sup 15}N and {sup 1}H assignments is given for all the amide NH and C{sup {alpha}}H resonances of all non-proline residues, as well as the {sup 1}H assignments for some of the amino acid side chains. This first example of the sequence-specific assignment of a protein using heteronuclear 3D NMR provides a basis for further conformational and dynamic studies of interleukin 1{beta}.

• OSTI ID: 6170376
• Journal Information: Biochemistry; (USA), Journal Name: Biochemistry; (USA) Vol. 29:14; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550601* -- Medicine-- Unsealed Radionuclides in Diagnostics
62 RADIOLOGY AND NUCLEAR MEDICINE
BARYONS
CHEMICAL SHIFT
DOUBLE LABELLING
ELEMENTARY PARTICLES
FERMIONS
GROWTH FACTORS
HADRONS
ISOTOPES
LABELLING
LIGHT NUCLEI
LYMPHOKINES
MAGNETIC RESONANCE
MITOGENS
NITROGEN 15
NITROGEN ISOTOPES
NUCLEAR MAGNETIC RESONANCE
NUCLEI
NUCLEONS
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
OVERHAUSER EFFECT
PEPTIDES
POLYPEPTIDES
PROTEINS
PROTONS
RESONANCE
STABLE ISOTOPES