Application of high resolution NMR, ESR, and gamma-ray scintillation spectroscopy to the study of ligand binding in proteins. [Torpedo californica]
Electron spin resonance spectroscopy has been employed to study the nature of the ligand binding site of alpha-1-antitrypsin. Spectra of spin-labeled alpha-1-antitrypsin were recorded at pH's ranging from 2.4 to 12.5. This data demonstrates the tight binding of the spin-label to the protease, and the sensitivity of the bound spin-label to informational changes in the protease inhibitor. A molecular dipstick approach has also been applied to this system and has yielded information on the geometry of the cleft accommodating the spin-label. /sup 160/Terbium(III) exchange experiments have been performed on the acetylcholine receptor protein isolated from Torpedo californica, employing a specially designed flow dialysis apparatus constructed in the laboratory. The apparatus is designed to allow continuous monitoring of /sup 160/Tb(III) gamma-ray emission from the protein compartment of the flow dialysis cell. Nicotinic ligand-induced displacement of /sup 160/Tb(III) from the nicotinic binding site of the receptor was monitored as a funtion of (1) the concentration of nicotinic ligand in the washout buffer, and (2) the nature of the nicotinic ligand in the buffer. Measured /sup 160/Tb(III) exchange half-lives indicate (1) a direct relationship between /sup 160/Tb(III) displacement and nicotinic ligand concentration in the wash-out buffer, and (2) an enhanced /sup 160/Tb(III) displacement for nicotinic agents possessing quaternary ammonium functions.
- Research Organization:
- Clark Univ., Worcester, MA (USA)
- OSTI ID:
- 5721257
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
PROTEINS
BIOCHEMICAL REACTION KINETICS
ELECTRON SPIN RESONANCE
EXPERIMENTAL DATA
FISHES
GAMMA SPECTROSCOPY
MEASURING METHODS
PEPTIDE HYDROLASES
TERBIUM 160
ANIMALS
AQUATIC ORGANISMS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
DATA
DAYS LIVING RADIOISOTOPES
ENZYMES
HYDROLASES
INFORMATION
INTERMEDIATE MASS NUCLEI
ISOTOPES
KINETICS
MAGNETIC RESONANCE
NUCLEI
NUMERICAL DATA
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
RADIOISOTOPES
RARE EARTH ISOTOPES
RARE EARTH NUCLEI
REACTION KINETICS
RESONANCE
SPECTROSCOPY
TERBIUM ISOTOPES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques