1OF1

KINETICS AND CRYSTAL STRUCTURE OF THE HERPES SIMPLEX VIRUS TYPE 1 THYMIDINE KINASE INTERACTING WITH (SOUTH)-METHANOCARBA-THYMIDINE

Summary for 1OF1

• Entry DOI: 10.2210/pdb1of1/pdb
• Related: 1E2H 1E2I 1E2J 1E2K 1E2L 1E2M 1E2N 1E2P 1KI2 1KI3 1KI4 1KI6 1KI7 1KI8 1QHI 1VTK 2KI5 3VTK
• Descriptor: THYMIDINE KINASE, (SOUTH)-METHANOCARBA-THYMIDINE, SULFATE ION, ... (4 entities in total)
• Functional Keywords: transferase, thymidine kinase, antiviral drug, enzyme-prodrug gene, kinase, dna synthesis, atp-binding
• Biological source: HERPES SIMPLEX VIRUS (TYPE 1 / STRAIN 17)
• Total number of polymer chains: 2
• Total formula weight: 82732.41

Authors

Claus, M.T.,Schelling, P.,Folkers, G.,Marquez, V.E.,Scapozza, L.,Schulz, G.E. (deposition date: 2003-04-03, release date: 2004-06-03, Last modification date: 2023-12-13)

Primary citation

Schelling, P.,Claus, M.T.,Johner, R.,Marquez, V.E.,Schulz, G.E.,Scapozza, L.
Biochemical and Structural Characterization of (South)-Methanocarbathymidine that Specifically Inhibits Growth of Herpes Simplex Virus Type 1 Thymidine Kinase-Transduced Osteosarcoma Cells
J.Biol.Chem., 279:32832-, 2004

PubMed Abstract: Two analogs of the natural nucleoside dT featuring a pseudosugar with fixed conformation in place of the deoxyribosyl residue (carbathymidine analogs) were biochemically and structurally characterized for their acceptance by both human cytosolic thymidine kinase isoenzyme 1 (hTK1) and herpes simplex virus type 1 thymidine kinase (HSV1 TK) and subsequently tested in cell proliferation assays. 3'-exo-Methanocarbathymidine ((South)-methanocarbathymidine (S)-MCT), which is a substrate for HSV1 TK, specifically inhibited growth of HSV1 TK-transduced human osteosarcoma cells with an IC(50) value in the range of 15 microM without significant toxicity toward both hTK1-negative (TK(-)) and non-transduced cells. 2'-exo-Methanocarbathymidine ((North)-methanocarbathymidine (N)-MCT), which is a weak substrate for hTK1 and a substantial one for HSV1 TK, induced a specific growth inhibition in HSV1 TK-transfected cells comparable to that of (S)-MCT and ganciclovir. A growth inhibition activity was also observed with (N)-MCT and ganciclovir in non-transduced cells in a cell line-dependent manner, whereas TK(-) cells were not affected. The presented 1.95-A crystal structure of the complex (S)-MCT.HSV1 TK explains both the more favorable binding affinity and catalytic turnover of (S)-MCT for HSV1 TK over the North analog. Additionally the plasticity of the active site of the enzyme is addressed by comparing the binding of (North)- and (South)-carbathymidine analogs. The presented study of these two potent candidate prodrugs for HSV1 TK gene-directed enzyme prodrug therapy suggests that (S)-MCT may be even safer to use than its North counterpart (N)-MCT.

PubMed: 15163659
DOI: 10.1074/JBC.M313343200

Experimental method

X-RAY DIFFRACTION (1.95 Å)