Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1978 Mar;5(3):975–985. doi: 10.1093/nar/5.3.975

Yeast seryl tRNA synthetase: interactions between the ATP binding site and the sites for tRNASer and L-serine.

U Pachmann, H G Zachau
PMCID: PMC342037  PMID: 417297

Abstract

T1 ribonuclease digestion of yeast tRNASer in the presence of seryl tRNA synthetase was used for monitoring the relationship between the substrate binding sites on the synthetase. It was found that (a) ATP displaces the tRNA from the synthetase with an effector affinity constant corresponding to the Km for ATP of 10 micron; (b) AMP and a number of nucleoside triphosphates, while influencing the rate of aminoacylation, do not displace the tRNA from the enzyme; (c) ADP and PPi inhibit the aminoacylation and the binding of tRNASer; (d) adenylyl diphosphonate is bound to the synthetase and lowers the protection of the tRNA against the nuclease attack in a similar way as does ATP; (e) interactions between the sites of L-serine and tRNASer could only be shown when both sites for serine were saturated and, in addition, the ATP analog or ADP was present. It is concluded that in seryl tRNA synthetase binding sites for ATP interact with the ones for tRNA as well as with the ones for serine. These findings contribute to the understanding of the mechanism of aminoacylation.

Full text

PDF
975

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bartmann P., Hanke T., Holler E. L-phenylalanine:tRNA ligase of Escherichia coli K10. A rapid kinetic investigation of the catalytic reaction. Biochemistry. 1975 Nov 4;14(22):4777–4786. doi: 10.1021/bi00693a001. [DOI] [PubMed] [Google Scholar]
  2. Cornish-Bowden A., Koshland D. E., Jr Diagnostic uses of the Hill (Logit and Nernst) plots. J Mol Biol. 1975 Jun 25;95(2):201–212. doi: 10.1016/0022-2836(75)90390-3. [DOI] [PubMed] [Google Scholar]
  3. Fittler F., Zachau H. G. Half molecules of serine-specific transfer ribonucleic acids from yeast. Arch Biochem Biophys. 1973 Apr;155(2):368–380. doi: 10.1016/0003-9861(73)90126-4. [DOI] [PubMed] [Google Scholar]
  4. Heider H., Gottschalk E., Cramer F. Isolation and characterization of seryl-tRNA synthetase from yeast. Eur J Biochem. 1971 May 11;20(1):144–152. doi: 10.1111/j.1432-1033.1971.tb01372.x. [DOI] [PubMed] [Google Scholar]
  5. Hirsch R., Zachau H. G. Isolierung und Charakterisierung der Seryl- und Phenylalanyl-tRNA-Synthetase aus Hefe. Hoppe Seylers Z Physiol Chem. 1976 Apr;357(4):509–526. [PubMed] [Google Scholar]
  6. Hustedt H., Flossdorf J., Kula M. R. Interacting binding sites of isoleucyl-tRNA synthetase from Escherichia coli studied by equilibrium partition. Eur J Biochem. 1977 Mar 15;74(1):199–202. doi: 10.1111/j.1432-1033.1977.tb11381.x. [DOI] [PubMed] [Google Scholar]
  7. Hyafil F., Jacques Y., Fayat G., Fromant M., Dessen P., Blanquet S. Methionyl-tRNA synthetase from Escherichia coli: active stoichiometry and stopped-flow analysis of methionyl adenylate formaiton. Biochemistry. 1976 Aug 24;15(17):3678–3685. doi: 10.1021/bi00662a006. [DOI] [PubMed] [Google Scholar]
  8. Hörz W., Meyer D., Zachau H. G. Nuclease digestion of synthetase x tRNA complexes. Eur J Biochem. 1975 May 6;53(2):533–539. doi: 10.1111/j.1432-1033.1975.tb04096.x. [DOI] [PubMed] [Google Scholar]
  9. Hörz W., Zachau H. G. Complexes of aminoacyl-tRNA synthetases with tRNAs as studies by partial nuclease digestion. Eur J Biochem. 1973 Jan 3;32(1):1–14. doi: 10.1111/j.1432-1033.1973.tb02571.x. [DOI] [PubMed] [Google Scholar]
  10. Jacques Y., Blanquet S. Interrelation between transfer RNA and amino-acid-activating sites of methionyl transfer RNA synthetase from Escherichia coli. Eur J Biochem. 1977 Oct 3;79(2):433–441. doi: 10.1111/j.1432-1033.1977.tb11825.x. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Pachmann U., Cronvall E., Rigler R., Hirsch R., Wintermeyer W., Zachau H. G. On the specificity of interactions between transfer ribonucleic acids and aminoacyl-tRNA synthetases. Eur J Biochem. 1973 Nov 1;39(1):265–273. doi: 10.1111/j.1432-1033.1973.tb03123.x. [DOI] [PubMed] [Google Scholar]
  13. Pachmann U., Zachau H. G. Yeast seryl tRNA synthetase: two sets of substrate sites involved in aminoacylation. Nucleic Acids Res. 1978 Mar;5(3):961–973. doi: 10.1093/nar/5.3.961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Riesner D., Pingoud A., Boehme D., Peters F., Maass G. Distinct steps in the specific binding of tRNA to aminoacyl-tRNA synthetase. Temperature-jump studies on the serine-specific system from yeast and the tyrosine-specific system from Escherichia coli. Eur J Biochem. 1976 Sep;68(1):71–80. doi: 10.1111/j.1432-1033.1976.tb10765.x. [DOI] [PubMed] [Google Scholar]
  15. Rigler R., Cronvall E., Ehrenberg M., Pachmann U., Hirsch R., Zachau H. G. Interactions between aminoacyl tRNA synthetases, tRNAs, and fluorescent dyes. FEBS Lett. 1971 Nov 1;18(2):193–198. doi: 10.1016/0014-5793(71)80443-x. [DOI] [PubMed] [Google Scholar]
  16. Rigler R., Pachmann U., Hirsch R., Zachau H. G. On the interaction of seryl-tRNA synthetase with tRNA Ser. A contribution to the problem of synthetase-tRNA recognition. Eur J Biochem. 1976 May 17;65(1):307–315. doi: 10.1111/j.1432-1033.1976.tb10418.x. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES