Comparative thermodynamic study of the interaction of some antifolates with dihydrofolate reductase

Gilli, Robert; Sari, J.C.; Lopez, C.; Rimet, Odile; Briand, Claudette

doi:10.1016/0167-4838(90)90083-r

Cited by 13 publications

(6 citation statements)

References 20 publications

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“…Methotrexate represents a classical ptertidine analogue, and trimethoprim is a lipophilic 2,4-diaminopyrimidine analogue. Gulli et al [32] determined Ki values by calorimetry for MTX and TMP, using native bovine DHFR purified by affinity chromatography (Table 6). Both Ki values determined for recombinant bDHFR were similar to those reported by Gilli et al [32].…”

Section: Resultsmentioning

confidence: 99%

Recombinant bovine dihydrofolate reductase produced by mutagenesis and nested PCR of murine dihydrofolate reductase cDNA

Cody

Mao

Queener

2008

Protein Expression and Purification

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Recent reports of the slow-tight binding inhibition of bovine liver dihydrofolate reductase (bDHFR) in the presence of polyphenols isolated from green tea leaves has spurred renewed interest in the biochemical properties of bDHFR. Earlier studies were done with native bDHFR but in order to validate models of polyphenol binding to bDHFR, larger quntities of bDHFR are necessary to support structural studies. Bovine DHFR differs from its closest sequence homologue, murine DHFR, by 19 amino acids. To obtain the bDHFR cDNA, murineDHFR cDNA was transformed by a series of nested PCR reactions to reproduce the amino acid coding sequence for bovine DHFR. The bovine liver DHFR cDNA has an open reading frame of 561 base pairs encoding a protein of 187 amino acids that has a high level of conservation at the primary sequence level with other DHFR enzymes, and more so for the amino acid residues in the active site of the mammalian DHFR enzymes. Expression of the bovine DHFR cDNA in bacterial cells produced a stable recombinant protein with high enzymatic activity and kinetic properties similar to those previously reported for the native protein.

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Section: Resultsmentioning

confidence: 99%

Recombinant bovine dihydrofolate reductase produced by mutagenesis and nested PCR of murine dihydrofolate reductase cDNA

Cody

Mao

Queener

2008

Protein Expression and Purification

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“…Support for this supposition comes from the structural and functional evidence for positive cooperativity of the binding of these two ligands at adjacent sites in DHFR enzymes from both eukaryotic and prokaryotic organisms. [33][34][35] For example, a 100-fold increase in binding affinity of MTX (at 10°C) was reported when NADPH was bound to the bovine liver DHFR 36 (see Table 1).…”

Section: Multiligand Binding Eventsmentioning

confidence: 97%

“…Conditions employed were 50 mM HEPES (pH 7.5) and 100 mM NaCl, in a volume of 5 L. ⌺ ϭ sum of individual ⌬T m values for two ligands when present alone (i.e., MTX and NADPH) under identical conditions. b A K d for NADPH binding to bovine liver DHFR could not be found; however, the K d for the human enzyme was reported to be 40 nM by Schweitzer et al 33 c Gilli et al 36 These conditions were for bovine liver DHFR at pH 6.8 (0.1 M potassium phosphate buffer) at 10°C. d Cooperativity for NADPH ϩ and MTX binding to Escherichia coli DHFR.…”

Section: Multiligand Binding Applicationsmentioning

confidence: 98%

High-Density Miniaturized Thermal Shift Assays as a General Strategy for Drug Discovery

Pantoliano¹,

Petrella²,

Kwasnoski³

et al. 2001

J Biomol Screen

258

350

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“…Over time, our laboratory has gained considerable experience with ITC to characterize the thermodynamics of various protein/ligand complexes. As examples, we measured the thermodynamic parameters of the interaction between: cephalosporins and human serum albumin (Briand et al, 1982); methotrexate and some of its metabolites with dihydrofolate reductase (Gilli et al, 1988) and thymidylate synthase (Gilli et al, 1990); Ca 2þ , Mg 2þ , and calmodulin (Gilli et al, 1998); coumarins and gyrase fragment (Lafitte et al, 2002); vinca alkaloid and calmodulin (Makarov et al, 2007).…”

Section: Rationalementioning

confidence: 99%

Microtubule and MAPs

Devred¹,

Barbier²,

Lafitte³

et al. 2010

Methods in Cell Biology

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Microtubules are implicated in many essential cellular processes such as architecture, cell division, and intracellular traffic, due to their dynamic instability. This dynamicity is tightly regulated by microtubule-associated proteins (MAPs), such as tau and stathmin. Despite extensive studies motivated by their central role in physiological functions and pathological role in neurodegenerative diseases and cancer, the precise mechanisms of tau and stathmin binding to tubulin and their consequences on microtubule stability are still not fully understood. One of the most crucial points missing is a quantitative thermodynamic description of their interaction with tubulin/microtubules and of the tubulin complexes formed upon these interactions. In this chapter, we will focus on the use of analytical ultracentrifugation, isothermal titration calorimetry, and nuclear magnetic resonance-three powerful and complementary techniques in the field of MAP-tubulin/microtubule interactions, in addition to the spectrometric techniques and co-sedimentation approach. We will present the limits of these techniques to study this particular interaction and precautions that need to be taken during MAPs preparation. Understanding the molecular mechanisms that govern MAPs action on microtubular network will not only shed new light on the role of this crucial family of protein in the biology of the cell, but also hopefully open new paths to increase the therapeutic efficiency of microtubule-targeting drugs in cancers therapies and neurodegeneratives diseases prevention.

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Comparative thermodynamic study of the interaction of some antifolates with dihydrofolate reductase

Cited by 13 publications

References 20 publications

Recombinant bovine dihydrofolate reductase produced by mutagenesis and nested PCR of murine dihydrofolate reductase cDNA

Recombinant bovine dihydrofolate reductase produced by mutagenesis and nested PCR of murine dihydrofolate reductase cDNA

High-Density Miniaturized Thermal Shift Assays as a General Strategy for Drug Discovery

Microtubule and MAPs

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