Solution studies of staphylococcal nuclease H124L. 1. Backbone sup 1 H and sup 15 N resonances and secondary structure of the unligated enzyme as identified by three-dimensional NMR spectroscopy
- Univ. of Wisconsin, Madison (United States)
The backbone {sup 1}H and {sup 15}N resonances of unligated staphylococcal nuclease H124L (recombinant protein produced in Escherichia coli whose sequence is identical to the nuclease produced by the V8 strain of Staphylococcus aureus) have been assigned by three-dimensional (3D) {sup 1}H-{sup 15}N NOESY-HMQC NMR spectroscopy at 14.1 tesla. The protein sample used in this study was labeled uniformly with {sup 15}N to a level greater than 95% by growing the E. coli host on a medium containing (99% {sup 15}N)ammonium sulfate as the sole nitrogen source. The assignments include 82% of the backbone {sup 1}H{sup N} and {sup 1}H{sup {alpha}} resonances as well as the {sup 15}N resonances of non-proline residues. Secondary structural elements ({alpha}-helices, {beta}-sheets, reverse turns, and loops) were determined by analysis of patterns of NOE connectivities present in the 3D spectrum.
- OSTI ID:
- 5488849
- Journal Information:
- Biochemistry; (United States), Vol. 31:3; ISSN 0006-2960
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
NUCLEASES
NUCLEAR MAGNETIC RESONANCE
AQUEOUS SOLUTIONS
ESCHERICHIA COLI
NITROGEN 15
OVERHAUSER EFFECT
PROTONS
STAPHYLOCOCCUS
BACTERIA
BARYONS
DISPERSIONS
ELEMENTARY PARTICLES
ENZYMES
ESTERASES
FERMIONS
HADRONS
HYDROLASES
ISOTOPES
LIGHT NUCLEI
MAGNETIC RESONANCE
MICROORGANISMS
MIXTURES
NITROGEN ISOTOPES
NUCLEI
NUCLEONS
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PHOSPHODIESTERASES
PROTEINS
RESONANCE
SOLUTIONS
STABLE ISOTOPES
550201* - Biochemistry- Tracer Techniques