• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. Mar 1981; 145(3): 1374–1385.
PMCID: PMC217142

Adenylate energy charge in Escherichia coli CR341T28 and properties of heat-sensitive adenylate kinase.

Abstract

Escherichia coli strain CR341T28 will not grow at temperatures above 34 degrees C in liquid medium, and the adenylate kinase of this strain is heat sensitive. When a culture was shifted from a permissive (30 degrees C) to a nonpermissive (36 degrees C) temperature, the adenylate energy charge fell from 0.9 to 0.2, with a concurrent decrease in the number of viable cells and in the specific activity of adenylate kinase. When cultures of the temperature-sensitive strain were grown at temperatures above 30 degrees C, the adenylate energy charge, the specific activity of adenylate kinase, and the growth rate were lower than the corresponding parameters for the parental strain. By isotopic labeling of the adenine nucleotides in vivo, it was determined that increasing growth temperatures between 30 and 34 degrees C for the heat-sensitive strain resulted in a decrease in the adenosine triphosphate-to-adenosine monophosphate and adenosine triphosphate-to-adenosine diphosphate ratios. Between 26 and 30 degrees C the adenosine triphosphate-to-adenosine diphosphate ratio was essentially normal in the temperature-sensitive strain, but the adenosine triphosphate-to-adenosine diphosphate ratio was decreased. The adenylate ratios in the parental strain did not change between 30 and 34 degrees C. The adenylate kinase mass action ratio for each strain was essentially constant under all growth conditions. When assayed at 30 degrees C, the affinities of the enzyme from the mutant strain were somewhat lower than those of the parent adenylate kinase. The mutant enzyme also did not exhibit the substrate inhibition that was observed at high adenosine monophosphate concentrations with the parental enzyme. An increase in the assay temperature from 30 degrees to 40 degrees C had little or no effect on the Km values determined for the parental adenylate kinase, but caused the Km values determined for the mutant adenylate kinase to increase by a factor of two or more.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.8M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Atkinson DE. Regulation of enzyme function. Annu Rev Microbiol. 1969;23:47–68. [PubMed]
  • Ball WJ, Jr, Atkinson DE. Adenylate energy charge in Saccharomyces cerevisiae during starvation. J Bacteriol. 1975 Mar;121(3):975–982. [PMC free article] [PubMed]
  • Chapman AG, Atkinson DE. Adenine nucleotide concentrations and turnover rates. Their correlation with biological activity in bacteria and yeast. Adv Microb Physiol. 1977;15:253–306. [PubMed]
  • Chapman AG, Fall L, Atkinson DE. Adenylate energy charge in Escherichia coli during growth and starvation. J Bacteriol. 1971 Dec;108(3):1072–1086. [PMC free article] [PubMed]
  • Cousin D. Mutants thermosensibles d'Escherichia coli K12. II. Etude d'une mutation létale affectant une réaction génératrice d'énercie. Ann Inst Pasteur (Paris) 1967 Sep;113(3):309–325. [PubMed]
  • Cousin D, Belaïch JP. Sur une mutatio thermosensible d'Escherichia coli affectant une fonction énergétique. C R Acad Sci Hebd Seances Acad Sci D. 1966 Sep 19;263(12):886–888. [PubMed]
  • Cousin D, Buttin G. Mutants thermosensibles d'Escherichia coli K12. 3. Une mutation létale d'E. coli affectant l'activité de l'adénylate-kinase. Ann Inst Pasteur (Paris) 1969 Nov;117(5):612–630. [PubMed]
  • Cronan JE., Jr Molecular biology of bacterial membrane lipids. Annu Rev Biochem. 1978;47:163–189. [PubMed]
  • Cronan JE, Jr, Ray TK, Vagelos PR. Selection and characterization of an E. coli mutant defective in membrane lipid biosynthesis. Proc Natl Acad Sci U S A. 1970 Mar;65(3):737–744. [PMC free article] [PubMed]
  • Glaser M, Nulty W, Vagelos PR. Role of adenylate kinase in the regulation of macromolecular biosynthesis in a putative mutant of Escherichia coli defective in membrane phospholipid biosynthesis. J Bacteriol. 1975 Jul;123(1):128–136. [PMC free article] [PubMed]
  • Goldberg AL, St John AC. Intracellular protein degradation in mammalian and bacterial cells: Part 2. Annu Rev Biochem. 1976;45:747–803. [PubMed]
  • Hochstadt-Ozer J, Cashel M. The regulation of purine utilization in bacteria. V. Inhibition of purine phosphoribosyltransferase activities and purine uptake in isolated membrane vesicles by guanosine tetraphosphate. J Biol Chem. 1972 Nov 10;247(21):7067–7072. [PubMed]
  • Holmes RK, Singer MF. Purification and characterization of adenylate kinase as an apparent adenosine triphosphate-dependent inhibitor of ribonuclease II in Escherichia coli. J Biol Chem. 1973 Mar 25;248(6):2014–2021. [PubMed]
  • Holms WH, Hamilton ID, Robertson AG. The rate of turnover of the adenosine triphosphate pool of Escherichia coli growing aerobically in simple defined media. Arch Mikrobiol. 1972;83(2):95–109. [PubMed]
  • Kohiyama M, Cousin D, Ryter A, Jacob F. Mutants thermosensibles d'Escherichia coli K 12. I. Isolement et caractérisation rapide. Ann Inst Pasteur (Paris) 1966 Apr;110(4):465–486. [PubMed]
  • Luria SE, Suit JL, Plate CA. Initiation of transcription is temperature-dependent in an E. coli mutant with ts adenylate kinase. Biochem Biophys Res Commun. 1975 Nov 3;67(1):353–358. [PubMed]
  • OLIVER IT, PEEL JL. Myokinase activity in microorganisms. Biochim Biophys Acta. 1956 May;20(2):390–392. [PubMed]
  • Schaffner W, Weissmann C. A rapid, sensitive, and specific method for the determination of protein in dilute solution. Anal Biochem. 1973 Dec;56(2):502–514. [PubMed]
  • Swedes JS, Dial ME, McLaughlin CS. Regulation of protein synthesis during early limitation of Saccharomyces cerevisiae. J Bacteriol. 1979 Apr;138(1):162–170. [PMC free article] [PubMed]
  • Swedes JS, Sedo RJ, Atkinson DE. Relation of growth and protein synthesis to the adenylate energy charge in an adenine-requiring mutant of Escherichia coli. J Biol Chem. 1975 Sep 10;250(17):6930–6938. [PubMed]
  • Thompson ST, Cass KH, Stellwagen E. Blue dextran-sepharose: an affinity column for the dinucleotide fold in proteins. Proc Natl Acad Sci U S A. 1975 Feb;72(2):669–672. [PMC free article] [PubMed]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...