High-level 2H/13C/15N labeling of proteins for NMR studies
| Autor: | Chih Chin Huang, Carol A. Fierke, Ronald A. Venters, Leonard D. Spicer, Ronald Trolard, Bennett T. Farmer |
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| Rok vydání: | 1995 |
| Předmět: |
Deuterium NMR
Carbon Isotopes Magnetic Resonance Spectroscopy Molecular Structure Nitrogen Isotopes Chemistry Stereochemistry Carbonic anhydrase II Proteins Pulse sequence Deuterium Biochemistry Mass Spectrometry chemistry.chemical_compound Nuclear magnetic resonance Isotopic shift Escherichia coli Humans Molecule Ammonium chloride Sodium acetate Spectroscopy Carbonic Anhydrases |
| Zdroj: | Journal of Biomolecular NMR. 5:339-344 |
| ISSN: | 1573-5001 0925-2738 |
| DOI: | 10.1007/bf00182275 |
| Popis: | The protein human carbonic anhydrase II (HCA II) has been isotopically labeled with 2H, 13C and 15N for high-resolution NMR assignment studies and pulse sequence development. To increase the sensitivity of several key 1H/13C/15N triple-resonance correlation experiments, 2H has been incorporated into HCA II in order to decrease the rates of 13C and 1HN T2 relaxation. NMR quantities of protein with essentially complete aliphatic 2H incorporation have been obtained by growth of E. coli in defined media containing D2O, [1,2-13C2, 99%] sodium acetate, and [15N, 99%] ammonium chloride. Complete aliphatic deuterium enrichment is optimal for 13C and 15N backbone NMR assignment studies, since the 13C and 1HN T2 relaxation times and, therefore, sensitivity are maximized. In addition, complete aliphatic deuteration increases both resolution and sensitivity by eliminating the differential 2H isotopic shift observed for partially deuterated CHnDm moieties. |
| Databáze: | OpenAIRE |
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