Abstract
The Escherichia coli purT encoded glycinamide ribonucleotide transformylase (GAR transformylase) serves as an alternate enzyme in the production of formyl GAR for use in de novo purine biosynthesis. This enzyme differs from the previously known purN encoded enzyme in size, sequence, and substrates; ATP and formate are required as opposed to formyl tetrahydrofolate. Kinetic studies of the wild-type PurT enzyme described here demonstrate that formyl phosphate behaves as a chemically and kinetically competent intermediate. The requirement for ATP and GAR in these reactions is consistent with previous steady-state kinetic results, which demonstrated that all substrates must be bound before catalysis. Kinetic characterization of a mutant, which releases formyl phosphate into solution, and positional isotope exchange studies also support the assignment of formyl phosphate as a plausible intermediate.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 6709-6716 |
| Number of pages | 8 |
| Journal | Biochemistry |
| Volume | 36 |
| Issue number | 22 |
| DOIs | |
| State | Published - Jun 3 1997 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
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Formyl phosphate : A proposed intermediate in the reaction catalyzed by Escherichia coli PurT GAR transformylase. / Marolewski, Ariane E.; Mattia, Karen M.; Warren, Mark S.; Benkovic, Stephen J.
In: Biochemistry, Vol. 36, No. 22, 03.06.1997, p. 6709-6716.Research output: Contribution to journal › Article
TY - JOUR
T1 - Formyl phosphate
T2 - A proposed intermediate in the reaction catalyzed by Escherichia coli PurT GAR transformylase
AU - Marolewski, Ariane E.
AU - Mattia, Karen M.
AU - Warren, Mark S.
AU - Benkovic, Stephen J.
PY - 1997/6/3
Y1 - 1997/6/3
N2 - The Escherichia coli purT encoded glycinamide ribonucleotide transformylase (GAR transformylase) serves as an alternate enzyme in the production of formyl GAR for use in de novo purine biosynthesis. This enzyme differs from the previously known purN encoded enzyme in size, sequence, and substrates; ATP and formate are required as opposed to formyl tetrahydrofolate. Kinetic studies of the wild-type PurT enzyme described here demonstrate that formyl phosphate behaves as a chemically and kinetically competent intermediate. The requirement for ATP and GAR in these reactions is consistent with previous steady-state kinetic results, which demonstrated that all substrates must be bound before catalysis. Kinetic characterization of a mutant, which releases formyl phosphate into solution, and positional isotope exchange studies also support the assignment of formyl phosphate as a plausible intermediate.
AB - The Escherichia coli purT encoded glycinamide ribonucleotide transformylase (GAR transformylase) serves as an alternate enzyme in the production of formyl GAR for use in de novo purine biosynthesis. This enzyme differs from the previously known purN encoded enzyme in size, sequence, and substrates; ATP and formate are required as opposed to formyl tetrahydrofolate. Kinetic studies of the wild-type PurT enzyme described here demonstrate that formyl phosphate behaves as a chemically and kinetically competent intermediate. The requirement for ATP and GAR in these reactions is consistent with previous steady-state kinetic results, which demonstrated that all substrates must be bound before catalysis. Kinetic characterization of a mutant, which releases formyl phosphate into solution, and positional isotope exchange studies also support the assignment of formyl phosphate as a plausible intermediate.
UR - http://www.scopus.com/inward/record.url?scp=0030904155&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0030904155&partnerID=8YFLogxK
U2 - 10.1021/bi962961p
DO - 10.1021/bi962961p
M3 - Article
C2 - 9184151
AN - SCOPUS:0030904155
VL - 36
SP - 6709
EP - 6716
JO - Biochemistry
JF - Biochemistry
SN - 0006-2960
IS - 22
ER -