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Titolo:
HIGH-LEVEL H-2 C-13/N-15 LABELING OF PROTEINS FOR NMR-STUDIES/
Autore:
VENTERS RA; HUANG CC; FARMER BT; TROLARD R; SPICER LD; FIERKE CA;
Indirizzi:
DUKE UNIV,MED CTR,DEPT BIOCHEM DURHAM NC 27710 BRISTOL MYERS SQUIBB PHARMACEUT RES INST,MACROMOLEC NMR PRINCETON NJ 08543 CAMBRIDGE ISOTOPE LABS ANDOVER MA 01810 DUKE UNIV,MED CTR,DEPT RADIOL DURHAM NC 27710
Titolo Testata:
Journal of biomolecular NMR
fascicolo: 4, volume: 5, anno: 1995,
pagine: 339 - 344
SICI:
0925-2738(1995)5:4<339:HHCLOP>2.0.ZU;2-E
Fonte:
ISI
Lingua:
ENG
Soggetto:
CARBONIC ANHYDRASE-II; LARGER PROTEINS; RNA-POLYMERASE; C-13; EXPRESSION; SPECTRA; H-1;
Keywords:
DEUTERIUM ISOTOPE LABELING; 3D HETERONUCLEAR NMR; HUMAN CARBONIC ANHYDRASE II;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
25
Recensione:
Indirizzi per estratti:
Citazione:
R.A. Venters et al., "HIGH-LEVEL H-2 C-13/N-15 LABELING OF PROTEINS FOR NMR-STUDIES/", Journal of biomolecular NMR, 5(4), 1995, pp. 339-344

Abstract

The protein human carbonic anhydrase II (HCA II) has been isotopically labeled with H-2, C-13 and N-15 for high-resolution NMR assignment studies and pulse sequence development. To increase the sensitivity of several key H-1/C-13/N-15 triple-resonance correlation experiments, H-2 has been incorporated into HCA II in order to decrease the rates of C-13 and H-1(N) T-2 relaxation. NMR quantities of protein with essentially complete aliphatic H-2 incorporation have been obtained by growthof E. coli in defined media containing D2O, [1,2-C-13(2), 99%] sodiumacetate, and [N-15, 99%] ammonium chloride. Complete aliphatic deuterium enrichment is optimal for C-13 and N-15 backbone NMR assignment studies, since the C-13 and H-1(N) T-2 relaxation times and, therefore, sensitivity are maximized. In addition, complete aliphatic deuterationincreases both resolution and sensitivity by eliminating the differential H-2 isotopic shift observed for partially deuterated CHnDm moieties.

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Documento generato il 22/09/20 alle ore 04:15:08