PMID- 10585460
OWN - NLM
STAT- MEDLINE
DCOM- 20000113
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 50
DP  - 1999 Dec 10
TI  - Aspartate 142 is involved in both hydrolase and dehydrogenase catalytic centers
      of 10-formyltetrahydrofolate dehydrogenase.
PG  - 35777-84
AB  - The enzyme 10-formyltetrahydrofolate dehydrogenase (FDH) catalyzes conversion of 
      10-formyltetrahydrofolate to tetrahydrofolate in either a dehydrogenase or
      hydrolase reaction. The hydrolase reaction occurs in a 310-residue amino-terminal
      domain of FDH (N(t)-FDH), whereas the dehydrogenase reaction requires the
      full-length enzyme. N(t)-FDH shares some sequence identity with several
      10-formyltetrahydrofolate-utilizing enzymes. All these enzymes have a strictly
      conserved aspartate, which is Asp(142) in the case of N(t)-FDH. Replacement of
      the aspartate with alanine, asparagine, glutamate, or glutamine in N(t)-FDH
      resulted in complete loss of hydrolase activity. All the mutants, however, were
      able to bind folate, although with lower affinity than wild-type N(t)-FDH. Six
      other aspartate residues located near the conserved Asp(142) were substituted
      with an alanine, and these substitutions did not result in any significant
      changes in the hydrolase activity. The expressed D142A mutant of the full-length 
      enzyme completely lost both hydrolase and dehydrogenase activities. This study
      shows that Asp(142) is an essential residue in the enzyme mechanism for both the 
      hydrolase and dehydrogenase reactions of FDH, suggesting that either the two
      catalytic centers of FDH are overlapped or the dehydrogenase reaction occurs
      within the hydrolase catalytic center.
FAU - Krupenko, S A
AU  - Krupenko SA
AD  - Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, 
      Tennessee 37232, USA. krupenko@musc.edu
FAU - Wagner, C
AU  - Wagner C
LA  - eng
GR  - DK15289/DK/NIDDK NIH HHS/United States
GR  - DK26657-19/DK/NIDDK NIH HHS/United States
GR  - DK46788/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Recombinant Proteins)
RN  - 30KYC7MIAI (Aspartic Acid)
RN  - EC 1.5.- (Oxidoreductases Acting on CH-NH Group Donors)
RN  - EC 1.5.1.6 (formyltetrahydrofolate dehydrogenase)
RN  - EC 3.5.- (Amidohydrolases)
SB  - IM
MH  - Amidohydrolases/chemistry/*metabolism
MH  - Amino Acid Sequence
MH  - Amino Acid Substitution
MH  - Animals
MH  - *Aspartic Acid
MH  - Catalytic Domain
MH  - Cell Line
MH  - Crystallography, X-Ray
MH  - Kinetics
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Oxidoreductases Acting on CH-NH Group Donors/*chemistry/*metabolism
MH  - Protein Conformation
MH  - Recombinant Proteins/chemistry/metabolism
MH  - Sequence Alignment
MH  - Spodoptera
MH  - Transfection
EDAT- 1999/12/10 00:00
MHDA- 1999/12/10 00:01
CRDT- 1999/12/10 00:00
PHST- 1999/12/10 00:00 [pubmed]
PHST- 1999/12/10 00:01 [medline]
PHST- 1999/12/10 00:00 [entrez]
AID - 10.1074/jbc.274.50.35777 [doi]
AID - S0021-9258(19)53311-7 [pii]
PST - ppublish
SO  - J Biol Chem. 1999 Dec 10;274(50):35777-84. doi: 10.1074/jbc.274.50.35777.