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  A designed equine herpes thymidine kinase (EHV4 TK) variant improves ganciclovir-induced cell-killing.

McSorley, T., Ort, S., Monnerjahn, C., & Konrad, M. (2014). A designed equine herpes thymidine kinase (EHV4 TK) variant improves ganciclovir-induced cell-killing. Biochemical Pharmacology, 87(3), 435-444. doi:10.1016/j.bcp.2013.11.011.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0015-8268-F Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0027-CD59-3
Genre: Journal Article

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 Creators:
McSorley, T., Author
Ort, S., Author
Monnerjahn, C., Author
Konrad, M.1, Author              
Affiliations:
1Research Group of Enzyme Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578612              

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Free keywords: Herpes simplex virus thymidine kinase; Equine herpes virus-4 thymidine kinase; Ganciclovir; Suicide gene therapy; Enzyme prodrug activation
 Abstract: The limitations of the ganciclovir (GCV)/herpes simplex virus thymidine kinase (HSV1 TK: EC 2.7.1.21) system as a suicide gene therapy approach have been extensively studied over the years. In our study, we focused on improving the cytotoxic profile of the GCV/equine herpes virus-4 thymidine kinase (EHV4 TK: EC 2.7.1.21) system. Our approach involved the structure-guided mutagenesis of EHV4 TK in order to switch its ability to preferentially phosphorylate the natural substrate deoxythymidine (dT) to that of GCV. We performed steady-state kinetic analysis, genetic complementation in a thymidine kinase-deficient Escherichia coli strain, isothermal titration calorimetry, and analysis of GCV-induced cell killing through generation of HEK 293 stable cell-lines expressing EHV4 TK mutants and wild-type EHV4 TK. We found that the EHV4 TK S144H-GFP mutant preferentially phosphorylates GCV and confers increased GCV-induced cytotoxicity compared to wild-type EHV4 TK.

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Language(s): eng - English
 Dates: 2013-12-042014-02-01
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1016/j.bcp.2013.11.011
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Title: Biochemical Pharmacology
Source Genre: Journal
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Pages: - Volume / Issue: 87 (3) Sequence Number: - Start / End Page: 435 - 444 Identifier: -