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Guanosine 5'-triphosphate (GTP) binding to Guinea pig liver transglutaminase modulates enzyme activity

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:5127024
;
The effect of guanine nucleotides on purified Guinea pig liver transglutaminase (Gpig TG) activity was investigated. Guanosine 5'-triphosphate (GTP) at concentrations 20-200 ..mu..M inhibited purified TG. GTP specifically inhibited Gpig TG, since platelet Factor XIIIa (platelet TG) was not inhibited by GTP. Kinetic studies showed that GTP was a reversible non-competitive inhibitor of Gpig TG (Ki = 90 ..mu..M). Ca/sup 2 +/ (5 mM) abolished GTP inhibition of TG. A GTP binding site on Gpig TG was demonstrated by several techniques. Gpig TG was bound specifically to a GTP-Agarose column, and binding was inhibited by Ca/sup 2 +/. GTP also inhibited trypsin proteolysis of Gpig TG when analyzed by SDS-PAGE. Ca/sup 2 +/ inhibited the effect of GTP on trypsin proteolysis of Gpig TG. GTP also specifically inhibited photoaffinity labelling of Gpig TG by 8-Azidoguanosine-5'-(..gamma..-/sup 32/P)triphosphate. Guanosine 5'-diphosphate (GDP) and the nonhydrolyzable GTP analog guanosine 5'-0-(3-thiotriphosphate) (GTP-..gamma..-S) also inhibited Gpig TG activity while guanosine-5'-monophosphate (GMP) and guanine free base had no effect. This is the first demonstration that guanine nucleotides play a role in regulating Gpig TG activity. In conclusion, they have demonstrated a GTP binding site on Gpig TG, which is modulated by Ca/sup 2 +/ and may function to regulate TG activity.
Research Organization:
Duke Univ., Durham, NC
OSTI ID:
5127024
Report Number(s):
CONF-8606151-
Conference Information:
Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 45:6
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AFFINITY
ALKALINE EARTH METAL COMPOUNDS
AMIDASES
ANIMALS
AROMATICS
AZAARENES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
CALCIUM COMPOUNDS
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
ENZYME ACTIVITY
ENZYMES
GLANDS
GUANOSINE
GUINEA PIGS
HETEROCYCLIC COMPOUNDS
HYDROLASES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LIGHT NUCLEI
LIVER
MAMMALS
MEMBRANE PROTEINS
NON-PEPTIDE C-N HYDROLASES
NUCLEI
NUCLEOSIDES
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
OXYGEN COMPOUNDS
PHOSPHATES
PHOSPHORUS 32
PHOSPHORUS COMPOUNDS
PHOSPHORUS ISOTOPES
PROTEINS
PURINES
RADIOISOTOPES
REACTION KINETICS
RECEPTORS
RIBOSIDES
RODENTS
TRACER TECHNIQUES
VERTEBRATES