Partial sup 1 H NMR assignments of the Escherichia coli dihydrofolate reductase complex with folate: Evidence for a unique conformation of bound folate
- Pennsylvania State Univ., Univ. Park (USA)
- Research Institute of Scripps Clinic, La Jolla, CA (USA)
Sequence-specific {sup 1}H assignments have been made for over 25% of the amino acid side chains of Escherichia coli dihydrofolate reductase complexed with folate by using a variety of two-dimensional techniques. Proton resonances were assigned by using a combination of site-directed mutagenesis and a knowledge of the X-ray crystal structure. Unique sets of NOE connectivities present in hydrophobic pockets were matched with the X-ray structure and used to assign many of the residues. Other residues, particularly those near or in the active site, were assigned by site-directed mutagenesis. The ability to assign unambiguosly the proton resonances of these catalytically important residues allowed for extensive networks of NOE connectivities to follow from these assignments. As a consequence of these assignments, the orientation of the pterin ring of folate could be determined, and its conformation is similar to that of the productive dihydrofolate complex. Under these experimental conditions, only one bound form of the pterin ring could be detected.
- OSTI ID:
- 5615021
- Journal Information:
- Biochemistry; (USA), Vol. 29:41; ISSN 0006-2960
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
FOLIC ACID
NUCLEAR MAGNETIC RESONANCE
OXIDOREDUCTASES
CHEMICAL SHIFT
ESCHERICHIA COLI
MUTAGENESIS
OVERHAUSER EFFECT
PROTONS
PTERIDINES
AMINO ACIDS
AROMATICS
AZAARENES
BACTERIA
BARYONS
CARBOXYLIC ACIDS
DRUGS
ELEMENTARY PARTICLES
ENZYMES
FERMIONS
HADRONS
HEMATINICS
HEMATOLOGIC AGENTS
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
MAGNETIC RESONANCE
MICROORGANISMS
NUCLEONS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
RESONANCE
VITAMIN B GROUP
VITAMINS
550201* - Biochemistry- Tracer Techniques