Phosphorylation-dephosphorylation of yeast pyruvate dehydrogenase
Pyruvate dehydrogenase complex (PDC) was purified to homogeneity from baker's yeast (Saccharomyces cerevisiae). No pyruvate dehydrogenase (PDH) kinase activity was detected at any stage of the purification. However, the purified PDC was phosphorylated and inactivated by purified PDH kinase from bovine kidney mitochondria, Mg/sup 2 +/, and (..gamma..-/sup 32/P)ATP. The protein-bound radioactivity was localized in the PDH ..cap alpha.. subunit. The phosphorylated, inactivated PDC was dephosphorylated and reactivated with purified bovine PDH phosphatase, Mg/sup 2 +/, and Ca/sup 2 +/. From a tryptic digest of phosphorylated yeast PDC a radioactive peptide was isolated by anion and reverse phase HPLC. The sequence of this tetradecapeptide is Tyr-Gly-Gly-His-Ser(P)-Met-Ser-Asp-Pro-Gly-Thr-Thr-Tyr-Arg. This sequence is very similar to the sequence of a tryptic phosphopeptide derived from the ..cap alpha.. subunit of bovine kidney and heart PDH: Tyr-His-Gly-His-Ser(P)-Met-Ser-Asp-Pro-Gly-Val-Ser-Tyr-Arg.
- Research Organization:
- Univ. of Texas, Austin
- OSTI ID:
- 7171411
- Report Number(s):
- CONF-8606151-; TRN: 86-039034
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:6; Conference: 76. annual meeting of the Federation of American Society for Experimental Biology, Washington, DC, USA, 8 Jun 1986
- Country of Publication:
- United States
- Language:
- English
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OXIDOREDUCTASES
PHOSPHORYLATION
PURIFICATION
PEPTIDES
AMINO ACID SEQUENCE
PHOSPHOTRANSFERASES
BIOCHEMICAL REACTION KINETICS
ATP
CALCIUM COMPOUNDS
CATIONS
CATTLE
KIDNEYS
LIQUID COLUMN CHROMATOGRAPHY
MAGNESIUM COMPOUNDS
MITOCHONDRIA
PHOSPHORUS 32
SACCHAROMYCES CEREVISIAE
TRACER TECHNIQUES
ALKALINE EARTH METAL COMPOUNDS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CELL CONSTITUENTS
CHARGED PARTICLES
CHEMICAL REACTIONS
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
DOMESTIC ANIMALS
ENZYMES
FUNGI
IONS
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LIGHT NUCLEI
MAMMALS
MICROORGANISMS
MOLECULAR STRUCTURE
NUCLEI
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANOIDS
ORGANS
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
PLANTS
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RUMINANTS
SACCHAROMYCES
SEPARATION PROCESSES
TRANSFERASES
VERTEBRATES
YEASTS
550201* - Biochemistry- Tracer Techniques