Overcoming the overlap problem in the assignment of sup 1 H NMR spectra of larger proteins by use of three-dimensional heteronuclear sup 1 H- sup 15 N Hartmann-Hahn-multiple quantum coherence and nuclear Overhauser-multiple quantum coherence spectroscopy: Application to interleukin 1. beta

Journal Article · Tue Jul 25 00:00:00 EDT 1989 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00441a004· OSTI ID:5442473

Marion, D; Driscoll, P C; Kay, L E; Wingfield, P T; Bax, A; Gronenborn, A M; Clore, G M ^[1]

---

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

The application of three-dimensional (3D) heteronuclear NMR spectroscopy to the sequential assignment of the {sup 1}H NMR spectra of larger proteins is presented, using uniformly labeled ({sup 15}N)interleukin 1{beta}, a protein of 153 residues and molecular mass of 17.4 kDa, as an example. The two-dimensional (2D) 600-MHz spectra of interleukin 1{beta} are too complex for complete analysis, owing to extensive cross-peak overlap and chemical shift degeneracy. The authors show that the combined use of 3D {sup 1}H-{sup 15}N Hartmann-Hahn-multiple quantum coherence (HOHAHA-HMQC) and nuclear Overhauser-multiple quantum coherence (NOESY-HMQC) spectroscopy, designed to provide the necessary through-bond and through-space correlations for sequential assignment, provides a practical general-purpose method for resolving ambiguities which severely limit the analysis of conventional 2D NMR spectra. The problem of amide NH chemical shift degeneracy in the {sup 1}H NMR spectrum is therefore effectively removed, and the assignment procedure simply involves inspecting a series of 2D {sup 1}H-{sup 1}H slices edited by the chemical shift of the directly bonded {sup 15}N atom. It is envisaged that the intrinsic simplicity of the 3D heteronuclear spectra, even for proteins of 150-200 residues, will permit the development of efficient computer-assisted or automated sequential assignment methods.

• OSTI ID: 5442473
• Journal Information: Biochemistry; (USA), Vol. 28:15; ISSN 0006-2960
• Country of Publication: United States
• Language: English

Related Subjects

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