Glycinamide Ribonucleotide Formyltransferase Inhibitor

2020 ◽  
Author(s):  
Keyword(s):  
Glycinamide Ribonucleotide Formyltransferase

Structural features of 5,10-dideaza-5,6,7,8-tetrahydrofolate that determine inhibition of mammalian glycinamide ribonucleotide formyltransferase

Biochemistry ◽  
1991 ◽  
Vol 30 (7) ◽  
pp. 1997-2006 ◽  
Author(s):  
Samuel W. Baldwin ◽  
Archie Tse ◽  
Lynn S. Gossett ◽  
Edward C. Taylor ◽  
Andre Rosowsky ◽  
...  
Keyword(s):  
Structural Features ◽  
Glycinamide Ribonucleotide Formyltransferase

scholarly journals Mapping the active site of the Haemophilus influenzae methionyl-tRNA formyltransferase: residues important for catalysis and tRNA binding

1999 ◽  
Vol 339 (1) ◽  
pp. 63-69 ◽  
Author(s):  
D. Trevor NEWTON ◽  
Dev MANGROO
Keyword(s):  
Haemophilus Influenzae ◽  
Active Site ◽  
Fluorescence Analysis ◽  
Basic Mechanism ◽  
Site Directed Mutagenesis ◽  
Active Site Residues ◽  
Trna Recognition ◽  
Glycinamide Ribonucleotide Formyltransferase ◽  
Essential Step ◽  
Trna Binding

Formylation of the initiator methionyl-tRNA by methionyl-tRNA formyltransferase (MTF) is an essential step in initiation of protein synthesis in eubacteria. Here, site-directed mutagenesis was used to identify active site residues of the Haemophilus influenzae MTF. Of the nine residues investigated, only Arg-41, Asn-107, His-109 and Asp-145 were important for the function of the H. influenzae MTF. Replacement of these residues with Ala resulted in a significant reduction in the efficiency of catalysis. Intrinsic fluorescence analysis indicated that this was not due to a defect in N10-formyltetrahydrofolate (fTHF) binding. The Asp-145 and Arg-41 mutations reduced the affinity of the enzyme for the initiator tRNA, whereas the Asn-107 and His-109 mutations affected catalysis but not tRNA binding. Replacement of Arg-41, His-109 and Asp-145 with functionally similar residues also affected the activity of the enzyme. The data suggest that Asn-107, His-109 and Asp-145 are catalytic residues, whereas Arg-41 is involved in tRNA recognition. In the Escherichia coli glycinamide ribonucleotide formyltransferase, which also uses fTHF as the formyl donor, Asn-106, His-108 and Asp-144 participate in the catalytic step. Together, these observations imply that this group of enzymes uses the same basic mechanism in formylating their substrates.

scholarly journals Correction to Structural and Enzymatic Analysis of Tumor-Targeted Antifolates That Inhibit Glycinamide Ribonucleotide Formyltransferase

Biochemistry ◽  
2017 ◽  
Vol 56 (7) ◽  
pp. 1025-1025
Author(s):  
Siobhan M. Deis ◽  
Arpit Doshi ◽  
Zhanjun Hou ◽  
Larry H. Matherly ◽  
Aleem Gangjee ◽  
...  
Keyword(s):  
Enzymatic Analysis ◽  
Glycinamide Ribonucleotide Formyltransferase

Thymidylate synthase gene expression in solid tumors predicts for response to pemetrexed in vitro

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 13058-13058 ◽  
Author(s):  
U. Eismann ◽  
O. Oberschmidt ◽  
M. Ehnert ◽  
J. Fleeth ◽  
F. Lüdtke ◽  
...  
Keyword(s):  
Gene Expression ◽  
Statistical Tests ◽  
Human Tumor ◽  
Soft Agar ◽  
Expression Data ◽  
Glycinamide Ribonucleotide Formyltransferase ◽  
Cloning Assay ◽  
Thymidilate Synthase ◽  
Soft Agar Cloning

13058 Background: Pemetrexed (P) is a novel antifolate which targets thymidilate synthase (TS), dihydrofolate reductase (DHFR), and glycinamide ribonucleotide formyltransferase (GARFT). The aim of the present study was to identify gene expression thresholds for these enzymes in human tumor specimens in order to separate P-sensitive from P-resistant patients. Methods: Soft-agar cloning assays were performed on freshly biopsied tumor cells exposed one hour to clinically achievable concentrations of P. In parallel, RNA was isolated, transcribed to cDNA and subsequently used for multiplex real-time PCR. Gene expression data were normalized against beta-actin transcripts followed by correlation against cloning assay results. Iterative calculations (fourfold analysis) were done for each enzyme separately to find the best cutoff for prediction of sensitivity to P. Results: Sensitive and resistant tumor samples were statistically significant different in gene expression of TS, DHFR, and GARFT (p < 0.003). 81% of all tumors with a TS copy number < 144 (related to 104 copies β-actin) were sensitive to P in vitro. (specificity = 0.69; chi2 = 14.14). Statistical tests demonstrated that gene expression of TS, DHFR, and GARFT are dependent variables and that TS transcription is the leading variable. The combination of TS, DHFR, and GARFT expression data was not superior to TS alone. Conclusions: TS expression is the most meaningful predictor for sensitivity (≤ 144 copies) or resistance (> 144 copies) to Pemetrexed in fresh tumor tissue. This observation forms a rationale for clinical trials using TS expression as predictor for clinical response. No significant financial relationships to disclose.

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