Abstract
Mutagenesis in vitro has been used to obtain mutant forms of the bifunctional enzyme, chorismate mutase/ prephenate dehydrogenase. Plasmid DNA containing the genes that code for the enzyme was treated with hydroxylamine and the resulting products were used to transform strains of Eschericliia coli. Two types of mutant were isolated. One contained enzyme which was niutase‐positive, dehydrogenase‐negative while the other not exhibit either activity. Kinetic and physical analysis of one of the purified monofunctional enzymes showed that the loss of dehydrogenase activity was due to modification of the binding site for NAD. The results open the way for molecular studies of structure‐function relationships with this bifunctional enzyme.
| Original language | English |
|---|---|
| Pages (from-to) | 513-519 |
| Number of pages | 7 |
| Journal | European Journal of Biochemistry |
| Volume | 124 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - 1 Jan 1982 |
| Externally published | Yes |
Cite this
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Characterization of Monofunctional Chorismate Mutase/Prephenate Dehydrogenase Enzymes Obtained via Mutagenesis of Recombinant Plasmids in vitro. / ROOD, Julian I.; PERROT, Brigitte; HEYDE, Elizabeth; MORRISON, John F.
In: European Journal of Biochemistry, Vol. 124, No. 3, 01.01.1982, p. 513-519.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - Characterization of Monofunctional Chorismate Mutase/Prephenate Dehydrogenase Enzymes Obtained via Mutagenesis of Recombinant Plasmids in vitro
AU - ROOD, Julian I.
AU - PERROT, Brigitte
AU - HEYDE, Elizabeth
AU - MORRISON, John F.
PY - 1982/1/1
Y1 - 1982/1/1
N2 - Mutagenesis in vitro has been used to obtain mutant forms of the bifunctional enzyme, chorismate mutase/ prephenate dehydrogenase. Plasmid DNA containing the genes that code for the enzyme was treated with hydroxylamine and the resulting products were used to transform strains of Eschericliia coli. Two types of mutant were isolated. One contained enzyme which was niutase‐positive, dehydrogenase‐negative while the other not exhibit either activity. Kinetic and physical analysis of one of the purified monofunctional enzymes showed that the loss of dehydrogenase activity was due to modification of the binding site for NAD. The results open the way for molecular studies of structure‐function relationships with this bifunctional enzyme.
AB - Mutagenesis in vitro has been used to obtain mutant forms of the bifunctional enzyme, chorismate mutase/ prephenate dehydrogenase. Plasmid DNA containing the genes that code for the enzyme was treated with hydroxylamine and the resulting products were used to transform strains of Eschericliia coli. Two types of mutant were isolated. One contained enzyme which was niutase‐positive, dehydrogenase‐negative while the other not exhibit either activity. Kinetic and physical analysis of one of the purified monofunctional enzymes showed that the loss of dehydrogenase activity was due to modification of the binding site for NAD. The results open the way for molecular studies of structure‐function relationships with this bifunctional enzyme.
UR - http://www.scopus.com/inward/record.url?scp=0019949352&partnerID=8YFLogxK
U2 - 10.1111/j.1432-1033.1982.tb06623.x
DO - 10.1111/j.1432-1033.1982.tb06623.x
M3 - Article
VL - 124
SP - 513
EP - 519
JO - European Journal of Biochemistry
JF - European Journal of Biochemistry
SN - 0014-2956
IS - 3
ER -