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

Title: (5'-/sup 32/P)-8-azidoguanosine-3',5'-monophosphate. I. Synthesis and properties. II. Interaction with E. coli proteins

Abstract

Under certain conditions of nutritional deprivation, microorganisms produce the magic spot nucleotides guanosine-3'-diphosphate-5'-triphosphate(pppGpp) and the tetraphosphate ppGpp. The latter is known to be a pleiotypic effector, i.e. it inhibits (and sometimes stimulates) many biological processes including transcription, translation, and metabolic pathways. It is unknown whether pppGpp, ppGp, pGpp, and pGp, other members of this family of guanosine-3',5'-phosphates, also have regulatory properties. To begin to investigate this question, a radioactive photoaffinity analog of pGp was prepared: (5'/sup 32/P)pN/sub 3/Gp. The interaction of this photoprobe with E. coli sonicates and a purified protein (RNA polymerase) was examined. At physiological salt concentrations two proteins (RNA polymerase) was examined. At physiological salt concentrations two proteins of 86,000 and 65,000 daltons (p86 and p65) were primarily photolabeled. Competition studies with guanosine and adenosine nucleotides indicated (5 /sup 32/P)pN/sub 3/Gp was labeling a ppGpp binding site on p86, and a pGp (or GMP) site on p65. ATP phosphorylation of p86 increased photoincorporation, while it decreased labeling of p65. The data also provide evidence of a different type of regulatory mechanism, i.e. phosphorylation modulates binding of an allosteric effector (ppGpp) to a protein(enzyme). Both ATP and GTP were found to phosphorylate the same proteins, although GTP wasmore » the preferred substrate in some cases.« less

Authors:
Publication Date:
Research Org.:
Wyoming Univ., Laramie (USA)
OSTI Identifier:
5873235
Resource Type:
Thesis/Dissertation
Resource Relation:
Other Information: Thesis (Ph. D.)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ESCHERICHIA COLI; BIOCHEMISTRY; GUANOSINE; BIOSYNTHESIS; CHEMICAL PROPERTIES; PROTEINS; PHOSPHORYLATION; ADENOSINE; ATP; CHEMICAL BONDS; LABELLED COMPOUNDS; NUCLEOTIDES; PHOSPHATES; PHOSPHORUS 32; PHOTOSENSITIVITY; TRACER TECHNIQUES; AROMATICS; AZAARENES; BACTERIA; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; CHEMICAL REACTIONS; CHEMISTRY; DAYS LIVING RADIOISOTOPES; HETEROCYCLIC COMPOUNDS; ISOTOPE APPLICATIONS; ISOTOPES; LIGHT NUCLEI; MICROORGANISMS; NUCLEI; NUCLEOSIDES; ODD-ODD NUCLEI; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; OXYGEN COMPOUNDS; PHOSPHORUS COMPOUNDS; PHOSPHORUS ISOTOPES; PURINES; RADIOISOTOPES; RIBOSIDES; SENSITIVITY; SYNTHESIS; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Owens, J.R. (5'-/sup 32/P)-8-azidoguanosine-3',5'-monophosphate. I. Synthesis and properties. II. Interaction with E. coli proteins. United States: N. p., 1983. Web.
Owens, J.R. (5'-/sup 32/P)-8-azidoguanosine-3',5'-monophosphate. I. Synthesis and properties. II. Interaction with E. coli proteins. United States.
Owens, J.R. Sat . "(5'-/sup 32/P)-8-azidoguanosine-3',5'-monophosphate. I. Synthesis and properties. II. Interaction with E. coli proteins". United States.
@article{osti_5873235,
title = {(5'-/sup 32/P)-8-azidoguanosine-3',5'-monophosphate. I. Synthesis and properties. II. Interaction with E. coli proteins},
author = {Owens, J.R.},
abstractNote = {Under certain conditions of nutritional deprivation, microorganisms produce the magic spot nucleotides guanosine-3'-diphosphate-5'-triphosphate(pppGpp) and the tetraphosphate ppGpp. The latter is known to be a pleiotypic effector, i.e. it inhibits (and sometimes stimulates) many biological processes including transcription, translation, and metabolic pathways. It is unknown whether pppGpp, ppGp, pGpp, and pGp, other members of this family of guanosine-3',5'-phosphates, also have regulatory properties. To begin to investigate this question, a radioactive photoaffinity analog of pGp was prepared: (5'/sup 32/P)pN/sub 3/Gp. The interaction of this photoprobe with E. coli sonicates and a purified protein (RNA polymerase) was examined. At physiological salt concentrations two proteins (RNA polymerase) was examined. At physiological salt concentrations two proteins of 86,000 and 65,000 daltons (p86 and p65) were primarily photolabeled. Competition studies with guanosine and adenosine nucleotides indicated (5 /sup 32/P)pN/sub 3/Gp was labeling a ppGpp binding site on p86, and a pGp (or GMP) site on p65. ATP phosphorylation of p86 increased photoincorporation, while it decreased labeling of p65. The data also provide evidence of a different type of regulatory mechanism, i.e. phosphorylation modulates binding of an allosteric effector (ppGpp) to a protein(enzyme). Both ATP and GTP were found to phosphorylate the same proteins, although GTP was the preferred substrate in some cases.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1983},
month = {1}
}

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: