Assignment of the backbone sup 1 H and sup 15 N NMR resonances of bacteriophage T4 lysozyme

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

DOI:https://doi.org/10.1021/bi00479a003· OSTI ID:6229192

McIntosh, L P; Dahlquist, F W ^[1]; Wand, A J ^[2]; Lowry, D F; Redfield, A G ^[3]

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1. Univ. of Oregon, Eugene (USA)
2. Fox Chase Cancer Center, Philadelphia, PA (USA)
3. Brandeis Univ., Waltham, MA (USA)

The proton and nitrogen ({sup 15}NH-H{sup {alpha}}-H{sup {beta}}) resonances of bacteriophage T4 lysozyme were assigned by {sup 15}N-aided {sup 1}H NMR. The assignments were directed from the backbone amide {sup 1}H-{sup 15}N nuclei, with the heteronuclear single-multiple-quantum coherence (HSMQC) spectrum of uniformly {sup 15}N enriched protein serving as the master template for this work. The main-chain amide {sup 1}H-{sup 15}N resonances and H{sup {alpha}} resonances were resolved and classified into 18 amino acid types by using HMQC and {sup 15}N-edited COSY measurements, respectively, of T4 lysozymes selectively enriched with one or more of {alpha}-{sup 15}N-labeled Ala, Arg, Asn, Asp, Gly, Gln, Glu, Ile, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val. The heteronuclear spectra were complemented by proton DQF-COSY and TOCSY spectra of unlabeled protein in H{sub 2}O and D{sub 2}O buffers, from which the H{sup {beta}} resonances of many residues were identified. The NOE cross peaks to almost every amide proton were resolved in {sup 15}N-edited NOESY spectra of the selectively {sup 15}N enriched protein samples. Residue specific assignments were determined by using NOE connectivities between protons in the {sup 15}NH-H{sup {alpha}}-H{sup {beta}} spin systems of known amino acid type. Additional assignments of the aromatic proton resonances were obtained from {sup 1}H NMR spectra of unlabeled and selectively deuterated protein samples. The secondary structure of T4 lysozyme indicated from a qualitative analysis of the NOESY data is consistent with the crystallographic model of the protein.

• OSTI ID: 6229192
• Journal Information: Biochemistry; (USA), Journal Name: Biochemistry; (USA) Vol. 29:27; 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
AMINO ACID SEQUENCE
AMMONIUM CHLORIDES
AMMONIUM COMPOUNDS
AMMONIUM HALIDES
BACTERIOPHAGES
BARYONS
CHEMICAL SHIFT
CHLORIDES
CHLORINE COMPOUNDS
ELEMENTARY PARTICLES
ENZYMES
FERMIONS
GLYCOSYL HYDROLASES
HADRONS
HALIDES
HALOGEN COMPOUNDS
HYDROLASES
ISOTOPES
LABELLED COMPOUNDS
LIGHT NUCLEI
LYSOZYME
MAGNETIC RESONANCE
MICROORGANISMS
MOLECULAR STRUCTURE
NITROGEN 15
NITROGEN ISOTOPES
NMR SPECTRA
NUCLEAR MAGNETIC RESONANCE
NUCLEI
NUCLEONS
O-GLYCOSYL HYDROLASES
ODD-EVEN NUCLEI
OVERHAUSER EFFECT
PARASITES
PROTONS
RESONANCE
SPECTRA
STABLE ISOTOPES
VIRUSES