Rational design in genetic engineering: dream or reality? A priori predictions for thrombin and ribonuclease a mutants

G. Náray-Szabó, Jenö Nagy, Attila Bérces

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Simple electrostatic potential calculations were made for thrombin and ribonuclease A in order to predict the effect of hypothetical point mutations on the catalytic rate, kcat. The numerical results were scaled using experimental data for trypsin and subtilisin mutants. The decrease in kcat in D102A and D102N mutants of thrombin was predicted to be five and four orders of magnitude as compared with the wild-type enzyme. Using the same scaling procedure, an increase in the catalytic rate of a factor of ten was predicted in the D121A mutant of ribonuclease A. On the other hand, in the K41A mutant of the same enzyme the estimated rate change was a decrease of two orders of magnitude. It was found that the effects of the mutations investigated depend almost exclusively on geometric factors. Accordingly, it is concluded that in all point mutants of serine proteases where the buried aspartate is replaced by alanine or asparagine, the rate decrease will vary between four and six orders of magnitude.

Original languageEnglish
Pages (from-to)401-412
Number of pages12
JournalJournal of Molecular Structure: THEOCHEM
Volume200
Issue numberC
DOIs
Publication statusPublished - 1989

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thrombin
Genetic engineering
Pancreatic Ribonuclease
Genetic Engineering
Ribonucleases
Thrombin
Enzymes
Subtilisin
Asparagine
Serine Proteases
mutations
predictions
Static Electricity
Point Mutation
Aspartic Acid
Alanine
Trypsin
enzymes
Electrostatics
aspartates

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Computational Theory and Mathematics
  • Atomic and Molecular Physics, and Optics

Cite this

Rational design in genetic engineering : dream or reality? A priori predictions for thrombin and ribonuclease a mutants. / Náray-Szabó, G.; Nagy, Jenö; Bérces, Attila.

In: Journal of Molecular Structure: THEOCHEM, Vol. 200, No. C, 1989, p. 401-412.

Research output: Contribution to journalArticle

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