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Titolo:
THE IDENTIFICATION OF METAL-BINDING LIGAND RESIDUES IN METALLOPROTEINS USING NUCLEAR-MAGNETIC-RESONANCE SPECTROSCOPY
Autore:
SCROFANI SDB; WRIGHT PE; DYSON HJ;
Indirizzi:
SCRIPPS RES INST,DEPT BIOL MOL,10550 N TORREY PINES RD LA JOLLA CA 92037 SCRIPPS RES INST,DEPT BIOL MOL LA JOLLA CA 92037 SCRIPPS RES INST,SKAGGS INST CHEM BIOL LA JOLLA CA 92037
Titolo Testata:
Protein science
fascicolo: 11, volume: 7, anno: 1998,
pagine: 2476 - 2479
SICI:
0961-8368(1998)7:11<2476:TIOMLR>2.0.ZU;2-R
Fonte:
ISI
Lingua:
ENG
Soggetto:
SIDE-CHAIN RESONANCES; BETA-LACTAMASE-II; BACTEROIDES-FRAGILIS; BACILLUS-CEREUS; BACKBONE AMIDE; NMR; PROTEINS; CD-113; CONNECTIVITIES; C-13;
Keywords:
METAL BINDING LIGANDS; METALLO-BETA-LACTAMASE; METALLOPROTEIN; NMR;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
18
Recensione:
Indirizzi per estratti:
Citazione:
S.D.B. Scrofani et al., "THE IDENTIFICATION OF METAL-BINDING LIGAND RESIDUES IN METALLOPROTEINS USING NUCLEAR-MAGNETIC-RESONANCE SPECTROSCOPY", Protein science, 7(11), 1998, pp. 2476-2479

Abstract

The identification of metal-binding ligands in metalloproteins is an important step in gaining detailed information regarding the environment of the active site. Traditionally, techniques such as Cd-113-substitution for the active metal followed by isotope-filtered NMR techniques have been used to this end. However, for medium to high molecular weight proteins (>20 kDa), these experiments may not be beneficial due to extensive H-1 spectral overlap. Here, we describe an alternative approach, where metal-binding ligands such as histidine and cysteine are specifically N-15 backbone labeled, excess EDTA is added and changes to {H-1-N-15} HSQC spectra are followed. Under these conditions, the amide groups of all N-15 labeled histidine and cysteine residues, which were either ligands or residues close to the active site, were identified unambiguously for metallo-beta-lactamase from Bacteroides fragilis.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 30/03/20 alle ore 19:44:59