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 language | English |
|---|---|
| Pages (from-to) | 401-412 |
| Number of pages | 12 |
| Journal | Journal of Molecular Structure: THEOCHEM |
| Volume | 200 |
| Issue number | C |
| DOIs | |
| Publication status | Published - 1989 |
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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 journal › Article
}
TY - JOUR
T1 - Rational design in genetic engineering
T2 - dream or reality? A priori predictions for thrombin and ribonuclease a mutants
AU - Náray-Szabó, G.
AU - Nagy, Jenö
AU - Bérces, Attila
PY - 1989
Y1 - 1989
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=26144458614&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=26144458614&partnerID=8YFLogxK
U2 - 10.1016/0166-1280(89)85068-7
DO - 10.1016/0166-1280(89)85068-7
M3 - Article
AN - SCOPUS:26144458614
VL - 200
SP - 401
EP - 412
JO - Computational and Theoretical Chemistry
JF - Computational and Theoretical Chemistry
SN - 2210-271X
IS - C
ER -