PMID- 12755606
OWN - NLM
STAT- MEDLINE
DCOM- 20030702
LR  - 20141120
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 42
IP  - 20
DP  - 2003 May 27
TI  - Rational design, synthesis, evaluation, and crystal structure of a potent 
      inhibitor of human GAR Tfase: 
      10-(trifluoroacetyl)-5,10-dideazaacyclic-5,6,7,8-tetrahydrofolic acid.
PG  - 6043-56
AB  - Glycinamide ribonucleotide transformylase (GAR Tfase) has been the target of 
      anti-neoplastic intervention for almost two decades. Here, we use a 
      structure-based approach to design a novel folate analogue, 
      10-(trifluoroacetyl)-5,10-dideazaacyclic-5,6,7,8-tetrahydrofolic acid 
      (10-CF(3)CO-DDACTHF, 1), which specifically inhibits recombinant human GAR Tfase 
      (K(i) = 15 nM), but is inactive (K(i) > 100 microM) against other 
      folate-dependent enzymes that have been examined. Moreover, compound 1 is a 
      potent inhibitor of tumor cell proliferation (IC(50) = 16 nM, CCRF-CEM), which 
      represents a 10-fold improvement over Lometrexol, a GAR Tfase inhibitor that has 
      been in clinical trials. Thus, this folate analogue 1 is among the most potent 
      and selective inhibitors known toward GAR Tfase. Contributing to its efficacious 
      activity, compound 1 is effectively transported into the cell by the reduced 
      folate carrier and intracellularly sequestered by polyglutamation. The crystal 
      structure of human GAR Tfase with folate analogue 1 at 1.98 A resolution 
      represents the first structure of any GAR Tfase to be determined with a cofactor 
      or cofactor analogue without the presence of substrate. The folate-binding loop 
      of residues 141-146, which is highly flexible in both Escherichia coli and 
      unliganded human GAR Tfase structures, becomes highly ordered upon binding 1 in 
      the folate-binding site. Computational docking of the natural cofactor into this 
      and other apo or complexed structures provides a rational basis for modeling how 
      the natural cofactor 10-formyltetrahydrofolic acid interacts with GAR Tfase, and 
      suggests that this folate analogue-bound conformation represents the best 
      template to date for inhibitor design.
FAU - Zhang, Yan
AU  - Zhang Y
AD  - Department of Molecular Biology, The Scripps Research Institute, 10550 North 
      Torrey Pines Road, La Jolla, California 92037, USA.
FAU - Desharnais, Joel
AU  - Desharnais J
FAU - Marsilje, Thomas H
AU  - Marsilje TH
FAU - Li, Chenglong
AU  - Li C
FAU - Hedrick, Michael P
AU  - Hedrick MP
FAU - Gooljarsingh, Lata T
AU  - Gooljarsingh LT
FAU - Tavassoli, Ali
AU  - Tavassoli A
FAU - Benkovic, Stephen J
AU  - Benkovic SJ
FAU - Olson, Arthur J
AU  - Olson AJ
FAU - Boger, Dale L
AU  - Boger DL
FAU - Wilson, Ian A
AU  - Wilson IA
LA  - eng
GR  - P01 CA63536/CA/NCI NIH HHS/United States
GR  - P41 RR08605/RR/NCRR NIH HHS/United States
GR  - R24 CA95830/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (10-(trifluoroacetyl)-5,10-dideazaacyclic-5,6,7,8-tetrahydrofolic acid)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Macromolecular Substances)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Tetrahydrofolates)
RN  - 6P3AVY8A7Q (lometrexol)
RN  - EC 2.1.2.- (Hydroxymethyl and Formyl Transferases)
RN  - EC 2.1.2.2 (Phosphoribosylglycinamide Formyltransferase)
SB  - IM
MH  - Binding Sites
MH  - Cell Line
MH  - Crystallography, X-Ray
MH  - Drug Design
MH  - Enzyme Inhibitors/chemical synthesis/*chemistry/*pharmacology
MH  - Escherichia coli/enzymology
MH  - Humans
MH  - Hydroxymethyl and Formyl Transferases/*antagonists & inhibitors/chemistry
MH  - In Vitro Techniques
MH  - Kinetics
MH  - Macromolecular Substances
MH  - Models, Molecular
MH  - Nuclear Magnetic Resonance, Biomolecular
MH  - Phosphoribosylglycinamide Formyltransferase
MH  - Protein Conformation
MH  - Recombinant Proteins/antagonists & inhibitors/chemistry
MH  - Static Electricity
MH  - Tetrahydrofolates/chemical synthesis/*chemistry/*pharmacology
EDAT- 2003/05/21 05:00
MHDA- 2003/07/03 05:00
CRDT- 2003/05/21 05:00
PHST- 2003/05/21 05:00 [pubmed]
PHST- 2003/07/03 05:00 [medline]
PHST- 2003/05/21 05:00 [entrez]
AID - 10.1021/bi034219c [doi]
PST - ppublish
SO  - Biochemistry. 2003 May 27;42(20):6043-56. doi: 10.1021/bi034219c.