Enzymic phosphoryl transfer to carbon and oxygen acceptors: An investigation of the biosynthesis of 2-aminoethylphosphonic acid in Tetrahymena pyriformis W. and the kinetic mechanism and cofactor controlled substrate specificity of yeast inorganic pyrophosphatase

Thesis/Dissertation · Thu Jan 01 00:00:00 EST 1987

OSTI ID:5518172

Barry, R J

This research is concerned with the study of two enzymatic systems which catalyze phosphoryl transfer reactions to carbon and oxygen acceptors. The first portion of this study is concerned with the elucidation of the T. pyriformis 2-aminoethylphosphonate (AEP) biosynthetic pathway. The de novo formation of AEP from exogenously added precursors in Tetrahymena cell-free preparations was evaluated by using radioisotopic techniques and NMR spectral analysis. Incubation of ({sup 32}P)-phosphoenolpyruvate (PEP) with cell-free preparations yielded ({sup 32}P)-labelled material that was chromatographically identical to authentic phosphonopyruvate (p-pry). A reexamination of AEP biosynthesis was initiated. In the second portion of this study the kinetic mechanism of yeast inorganic pyrophosphatase (PPase) was examined by carrying-out initial velocity studies. Ca{sup 2+} and Rh(H{sub 2}O){sub 4} (methylenediphosphonate) (Rh(H{sub 2}O){sub 4}PCP) were used as dead-end inhibitors to study the order of binding of Cr(H{sub 2}O){sub 4}PP to the substrate site and Mg{sup 2+} to the low affinity activator site on the enzyme.

• Research Organization: Maryland Univ., College Park, MD (USA)
• OSTI ID: 5518172
• Resource Relation: Other Information: Thesis (Ph. D.)
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
PHOSPHATASES
BIOCHEMICAL REACTION KINETICS
PHOSPHONIC ACID ESTERS
BIOSYNTHESIS
SUBSTRATES
SPECIFICITY
BIOLOGICAL PATHWAYS
CHROMATOGRAPHY
ENZYME INHIBITORS
MAGNESIUM COMPOUNDS
NMR SPECTRA
PHOSPHORUS 32
PRECURSOR
TETRAHYMENA
TRACER TECHNIQUES
YEASTS
ALKALINE EARTH METAL COMPOUNDS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CILIATA
DAYS LIVING RADIOISOTOPES
ENZYMES
ESTERASES
ESTERS
EUMYCOTA
FUNGI
HYDROLASES
INVERTEBRATES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LIGHT NUCLEI
MICROORGANISMS
NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
PHOSPHORUS ISOTOPES
PLANTS
PROTOZOA
RADIOISOTOPES
REACTION KINETICS
SEPARATION PROCESSES
SPECTRA
SYNTHESIS
550201* - Biochemistry- Tracer Techniques