Journal ArticleDOI
Kinetics and crystal structure of the wild-type and the engineered Y101F mutant of Herpes simplex virus type 1 thymidine kinase interacting with (North)-methanocarba-thymidine.
Andrea E. Prota,Joachim Vogt,Beatrice D. Pilger,Remo Perozzo,Christine Wurth,Victor E. Marquez,Pamela Russ,Georg E. Schulz,Gerd Folkers,Leonardo Scapozza +9 more
TLDR
TKHSV1(Y101F) shows a decrease in the quotient of the catalytic efficiency (kcat/KM) of dT over MCT corresponding to an increased specificity for MCT when compared to the wild-type enzyme.Abstract:
Kinetic and crystallographic analyses of wild-type Herpes simplex virus type 1 thymidine kinase (TKHSV1) and its Y101F-mutant [TKHSV1(Y101F)] acting on the potent antiviral drug 2‘-exo-methanocarba-thymidine (MCT) have been performed. The kinetic study reveals a 12-fold KM increase for thymidine processed with Y101F as compared to the wild-type TKHSV1. Furthermore, MCT is a substrate for both wild-type and mutant TKHSV1. Its binding affinity for TKHSV1 and TKHSV1(Y101F), expressed as Ki, is 11 μM and 51 μM, respectively, whereas the Ki for human cytosolic thymidine kinase is as high as 1.6 mM, rendering TKHSV1 a selectivity filter for antiviral activity. Moreover, TKHSV1(Y101F) shows a decrease in the quotient of the catalytic efficiency (kcat/KM) of dT over MCT corresponding to an increased specificity for MCT when compared to the wild-type enzyme. Crystal structures of wild-type and mutant TKHSV1 in complex with MCT have been determined to resolutions of 1.7 and 2.4 A, respectively. The thymine moiety o...read more
Citations
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Protein-based virtual screening of chemical databases. 1. Evaluation of different docking/scoring combinations.
TL;DR: A two-step protocol for screening large databases is proposed: screening of a reduced dataset containing a few known ligands for deriving the optimal docking/consensus scoring scheme and applying the latter parameters to the screening of the entire database.
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Structure and function of cellular deoxyribonucleoside kinases.
TL;DR: An overview of the structural studies made on this family of enzymes, including the catalytic mechanism as well as the mechanism for feedback inhibition, and homology models of other proteins in the family are made.
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Prodrugs for Gene-Directed Enzyme-Prodrug Therapy (Suicide Gene Therapy).
TL;DR: It is expected that continued studies to optimize the bystander effects and other properties of prodrugs and the activated species they generate will contribute to improvements in the effectiveness of suicide gene therapy.
Journal ArticleDOI
Characterization of CD20-Transduced T Lymphocytes as an Alternative Suicide Gene Therapy Approach for the Treatment of Graft-Versus-Host Disease
M Serafini,Massimiliano Manganini,Gianmaria Borleri,Martín Hernán Bonamino,L Imberti,Andrea Biondi,Josée Golay,Alessandro Rambaldi,Martino Introna +8 more
TL;DR: The CD20 transgene did not alter the functionality of T lymphocytes with respect to allogeneic recognition and cytotoxic response, anti-Epstein-Barr virus cytot toxic response, antigenic response to tetanus toxoid antigen, interleukin 2 (IL-2), IL-4, and interferon gamma production.
Journal ArticleDOI
Experimental and structural evidence that herpes 1 kinase and cellular DNA polymerase(s) discriminate on the basis of sugar pucker.
Victor E. Marquez,Tsipi Ben-Kasus,Joseph J. Barchi,Karen M. Green,Marc C. Nicklaus,Riad Agbaria +5 more
TL;DR: A combined solid-state and solution conformational analysis of both compounds, coupled with the direct measurement of mono-, di-, and triphosphate levels in control cells, cells infected with the Herpes simplex virus, or cells transfected with the corresponding viral kinase gene (HSV-tk), suggests that kinases prefer substrates that adopt the S sugar conformation.
References
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Journal ArticleDOI
PROCHECK: a program to check the stereochemical quality of protein structures
TL;DR: The PROCHECK suite of programs as mentioned in this paper provides a detailed check on the stereochemistry of a protein structure and provides an assessment of the overall quality of the structure as compared with well refined structures of the same resolution.
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MOLSCRIPT: a program to produce both detailed and schematic plots of protein structures
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Journal ArticleDOI
Nucleosides with a twist. Can fixed forms of sugar ring pucker influence biological activity in nucleosides and oligonucleotides
Victor E. Marquez,Maqbool A. Siddiqui,Abdallah Ezzitouni,Pamela Russ,Jianying Wang,Richard W. Wagner,Mark D. Matteucci +6 more
TL;DR: In this article, a bicyclo[310]hexane template was used to fix the ring pucker of 2'-deoxy-methanocarba-nucleosides to values corresponding to either one of these two extreme conformations that are typical of nucleosides.