scholarly journals Mg2+ Binds to the Surface of Thymidylate Synthase and Affects Hydride Transfer at the Interior Active Site

2013 ◽  
Vol 135 (20) ◽  
pp. 7583-7592 ◽  
Author(s):  
Zhen Wang ◽  
Paul J. Sapienza ◽  
Thelma Abeysinghe ◽  
Calvin Luzum ◽  
Andrew L. Lee ◽  
...  
Keyword(s):  
Thymidylate Synthase ◽  
Active Site ◽  
Hydride Transfer

scholarly journals The influence of active site conformations on the hydride transfer step of the thymidylate synthase reaction mechanism

2015 ◽  
Vol 17 (46) ◽  
pp. 30793-30804 ◽  
Author(s):  
Katarzyna Świderek ◽  
Amnon Kohen ◽  
Vicent Moliner
Keyword(s):  
Reaction Mechanism ◽  
Thymidylate Synthase ◽  
Active Site ◽  
Hydride Transfer ◽  
Md Simulations ◽  
Concerted Mechanism ◽  
X Ray ◽  
Rate Limiting Step ◽  
Limiting Step ◽  
Rate Limiting

QM/MM MD simulations from different X-ray structures support the concerted mechanism character in the rate limiting step of thymidylate synthase catalysis.

Tryptophan 80 and Leucine 143 Are Critical for the Hydride Transfer Step of Thymidylate Synthase by Controlling Active Site Access†,‡

Biochemistry ◽  
2002 ◽  
Vol 41 (22) ◽  
pp. 7021-7029 ◽  
Author(s):  
Timothy A. Fritz ◽  
Lu Liu ◽  
Janet S. Finer-Moore ◽  
Robert M. Stroud
Keyword(s):  
Thymidylate Synthase ◽  
Active Site ◽  
Hydride Transfer ◽  
Transfer Step ◽  
Site Access

scholarly journals An enzymatic activation of formaldehyde for nucleotide methylation

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Charles Bou-Nader ◽  
Frederick W. Stull ◽  
Ludovic Pecqueur ◽  
Philippe Simon ◽  
Vincent Guérineau ◽  
...  
Keyword(s):  
Amino Acids ◽  
Thymidylate Synthase ◽  
Active Site ◽  
De Novo ◽  
Crystallographic Structure ◽  
De Novo Synthesis ◽  
Active Site Residues ◽  
Enzymatic Activation ◽  
Enzyme Cofactors ◽  
Unique Ability

AbstractFolate enzyme cofactors and their derivatives have the unique ability to provide a single carbon unit at different oxidation levels for the de novo synthesis of amino-acids, purines, or thymidylate, an essential DNA nucleotide. How these cofactors mediate methylene transfer is not fully settled yet, particularly with regard to how the methylene is transferred to the methylene acceptor. Here, we uncovered that the bacterial thymidylate synthase ThyX, which relies on both folate and flavin for activity, can also use a formaldehyde-shunt to directly synthesize thymidylate. Combining biochemical, spectroscopic and anaerobic crystallographic analyses, we showed that formaldehyde reacts with the reduced flavin coenzyme to form a carbinolamine intermediate used by ThyX for dUMP methylation. The crystallographic structure of this intermediate reveals how ThyX activates formaldehyde and uses it, with the assistance of active site residues, to methylate dUMP. Our results reveal that carbinolamine species promote methylene transfer and suggest that the use of a CH2O-shunt may be relevant in several other important folate-dependent reactions.

Active site arginine controls the stereochemistry of hydride transfer in cyclohexanone monooxygenase

2018 ◽  
Vol 659 ◽  
pp. 47-56 ◽  
Author(s):  
Osei Boakye Fordwour ◽  
Kirsten R. Wolthers
Keyword(s):  
Active Site ◽  
Hydride Transfer ◽  
Cyclohexanone Monooxygenase

scholarly journals A Remote Mutation Affects the Hydride Transfer by Disrupting Concerted Protein Motions in Thymidylate Synthase

2012 ◽  
Vol 134 (42) ◽  
pp. 17722-17730 ◽  
Author(s):  
Zhen Wang ◽  
Thelma Abeysinghe ◽  
Janet S. Finer-Moore ◽  
Robert M. Stroud ◽  
Amnon Kohen
Keyword(s):  
Thymidylate Synthase ◽  
Hydride Transfer ◽  
Protein Motions

scholarly journals Crystal Structure of the FAD-Containing Ferredoxin-NADP+Reductase from the Plant PathogenXanthomonas axonopodispv. citri

2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
María Laura Tondo ◽  
Ramon Hurtado-Guerrero ◽  
Eduardo A. Ceccarelli ◽  
Milagros Medina ◽  
Elena G. Orellano ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Transfer Process ◽  
Plant Pathogen ◽  
Active Site ◽  
Functional Divergence ◽  
Hydride Transfer ◽  
Citrus Canker ◽  
Xanthomonas Axonopodis ◽  
C Terminus

We have solved the structure of ferredoxin-NADP(H) reductase, FPR, from the plant pathogenXanthomonas axonopodispv. citri, responsible for citrus canker, at a resolution of 1.5 Å. This structure reveals differences in the mobility of specific loops when compared to other FPRs, probably unrelated to the hydride transfer process, which contributes to explaining the structural and functional divergence between the subclass I FPRs. Interactions of the C-terminus of the enzyme with the phosphoadenosine of the cofactor FAD limit its mobility, thus affecting the entrance of nicotinamide into the active site. This structure opens the possibility of rationally designing drugs against theX. axonopodispv. citri phytopathogen.

scholarly journals Drugging Protein-Protein Interfaces of a Supramolecular Assembly as a Means to Overcome Resistance to Active Site Thymidylate Synthase Inhibitors

2020 ◽  
Vol 118 (3) ◽  
pp. 515a
Author(s):  
Tigran M. Abramyan ◽  
Alexander Tropsha ◽  
Andrew L. Lee ◽  
Paul J. Sapienza
Keyword(s):  
Thymidylate Synthase ◽  
Active Site ◽  
Supramolecular Assembly ◽  
Protein Interfaces ◽  
Thymidylate Synthase Inhibitors

scholarly journals Inter-Active Site Communication Mediated by the Dimer Interface β-Sheet in the Half-the-Sites Enzyme, Thymidylate Synthase

Biochemistry ◽  
2019 ◽  
Vol 58 (30) ◽  
pp. 3302-3313 ◽  
Author(s):  
Paul J. Sapienza ◽  
Konstantin I. Popov ◽  
David D. Mowrey ◽  
Bradley T. Falk ◽  
Nikolay V. Dokholyan ◽  
...  
Keyword(s):  
Thymidylate Synthase ◽  
Active Site ◽  
Dimer Interface ◽  
Β Sheet

scholarly journals Structural and Functional Characterization of the Human Thymidylate Synthase (hTS) Interface Variant R175C, New Perspectives for the Development of hTS Inhibitors

Molecules ◽  
2019 ◽  
Vol 24 (7) ◽  
pp. 1362
Author(s):  
Cecilia Pozzi ◽  
Stefania Ferrari ◽  
Rosaria Luciani ◽  
Maria Costi ◽  
Stefano Mangani
Keyword(s):  
Thymidylate Synthase ◽  
Active Site ◽  
Functional Characterization ◽  
Binding Mode ◽  
Anticancer Chemotherapy ◽  
Innovative Strategies ◽  
X Ray Crystallography ◽  
Site Targeting ◽  
New Strategies

Human thymidylate synthase (hTS) is pivotal for cell survival and proliferation, indeed it provides the only synthetic source of dTMP, required for DNA biosynthesis. hTS represents a validated target for anticancer chemotherapy. However, active site-targeting drugs towards hTS have limitations connected to the onset of resistance. Thus, new strategies have to be applied to effectively target hTS without inducing resistance in cancer cells. Here, we report the generation and the functional and structural characterization of a new hTS interface variant in which Arg175 is replaced by a cysteine. Arg175 is located at the interface of the hTS obligate homodimer and protrudes inside the active site of the partner subunit, in which it provides a fundamental contribution for substrate binding. Indeed, the R175C variant results catalytically inactive. The introduction of a cysteine at the dimer interface is functional for development of new hTS inhibitors through innovative strategies, such as the tethering approach. Structural analysis, performed through X-ray crystallography, has revealed that a cofactor derivative is entrapped inside the catalytic cavity of the hTS R175C variant. The peculiar binding mode of the cofactor analogue suggests new clues exploitable for the design of new hTS inhibitors.

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