Structural characterization of selenosubtilisin by sup 77 Se-NMR spectroscopy

Conference · Fri Mar 15 04:00:00 EST 1991 · FASEB Journal (Federation of American Societies for Experimental Biology); (United States)

OSTI ID:5217028

House, K L; Dunlap, R B; Odom, J D; Wu, Z P; Hilvert, D ^[1]

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1. Univ. of South Carolina, Columbia (United States) Research Inst. of Scripps Clinic, La Jolla, CA (United States)

Selenosubtilisin is an artificial enzyme containing an active site selenocysteine residue. In this environment the selenium atom is a valuable probe of structure-function relationships and also confers novel redox and hydrolytic properties to the original protease template. The authors have used {sup 77}Se NMR spectroscopy to characterize different oxidation states of {sup 77}Se isotopically enriched selenosubtilisin. The oxidized form of the enzyme exhibits a {sup 77}Se resonance at 1,189 ppm. This is in good agreement with the {sup 77}Se chemical shifts for model seleninic acids, confirming that the prosthetic group is in the seleninic acid oxidation state. On treatment of the oxidized enzyme with three equivalents of 3-carboxy-4-nitrobenzenethiol at pH 5.0, they observe the enzyme bound selenenyl sulfide at 388.5 ppm. This work demonstrates the utility of {sup 77}Se NMR spectroscopy for examining structure-function relationships of selenium containing proteins.

• OSTI ID: 5217028
• Report Number(s): CONF-9104107--
• Conference Information: Journal Name: FASEB Journal (Federation of American Societies for Experimental Biology); (United States) Journal Volume: 5:5
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
CHEMICAL SHIFT
ENZYMES
EVEN-ODD NUCLEI
INTERMEDIATE MASS NUCLEI
ISOMERIC TRANSITION ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
MAGNETIC RESONANCE
MOLECULAR STRUCTURE
NMR SPECTRA
NUCLEAR MAGNETIC RESONANCE
NUCLEI
ORGANIC COMPOUNDS
PROTEINS
RADIOISOTOPES
RESONANCE
SECONDS LIVING RADIOISOTOPES
SELENIUM 77
SELENIUM ISOTOPES
SPECTRA
STABLE ISOTOPES
STRUCTURE-ACTIVITY RELATIONSHIPS
TRACER TECHNIQUES