skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: 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

Abstract

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.

Authors:
Publication Date:
Research Org.:
Maryland Univ., College Park, MD (USA)
OSTI Identifier:
5518172
Resource Type:
Thesis/Dissertation
Resource Relation:
Other Information: Thesis (Ph. D.)
Country of Publication:
United States
Language:
English
Subject:
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

Citation Formats

Barry, R J. 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. United States: N. p., 1987. Web.
Barry, R J. 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. United States.
Barry, R J. 1987. "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". United States.
@article{osti_5518172,
title = {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},
author = {Barry, R J},
abstractNote = {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.},
doi = {},
url = {https://www.osti.gov/biblio/5518172}, journal = {},
number = ,
volume = ,
place = {United States},
year = {Thu Jan 01 00:00:00 EST 1987},
month = {Thu Jan 01 00:00:00 EST 1987}
}

Thesis/Dissertation:
Other availability
Please see Document Availability for additional information on obtaining the full-text document. Library patrons may search WorldCat to identify libraries that hold this thesis or dissertation.

Save / Share: