Zur Entwicklung eines Pharmakophormodells für thymidinkinaseabhängige, nukleosidanaloge Virostatika

Folkers, Gerd; Sakahara, Keiko; Schwobel, W; Eger, Kurt

doi:10.1002/ardp.19893220703

Cited by 5 publications

(2 citation statements)

References 10 publications

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“…*The model coordinates are available upon request from G. Folkers in mol or pdb file formats. Considerable support for the docking of the nucleosides into HSV1-TK comes from the fact that the relative spatial orientation of DHPG and thymidine is very similar to that found in an earlier study by active analogue approach [44], the 5-methyl-group of thymidine pointing in the direction of the N1 in the DHPG six-membered ring (Figs. 14 and 16).…”

Section: Resultsmentioning

confidence: 67%

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Computer-aided active-site-directed modeling of the Herpes Simplex Virus 1 and human thymidine kinase

Folkers

Trumpp-Kallmeyer

Gutbrod

et al. 1991

J Computer-Aided Mol Des

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Thymidine kinase (TK), which is induced by Herpes Simplex Virus 1 (HSV1), plays a key role in the antiviral activity of guanine derivatives such as aciclovir (ACV). In contrast, ACV shows only low affinity to the corresponding host cell enzyme. In order to define the differences in substrate binding of the two enzymes on molecular level, models for the three-dimensional (3-D) structures of the active sites of HSV1-TK and human TK were developed. The reconstruction of the active sites of HSV1-TK and human TK were developed. The reconstruction of the active sites started from primary and secondary structure analysis of various kinases. The results were validated to homologous enzymes with known 3-D structures. The models predict that both enzymes consist of a central core beta-sheet structure, connected by loops and alpha-helices very similar to the overall structure of other nucleotide binding enzymes. The phosphate binding site is made up of a highly conserved glycine-rich loop at the N-terminus of the proteins and a conserved region at the C-terminus. The thymidine recognition site was found about 100 amino acids downstream from the phosphate binding loop. The differing substrate specificity of human and HSV1-TK can be explained by amino-acid substitutions in the homologous regions. To achieve a better understanding of the structure of the active site and how the thymidine kinase proteins interact with their substrates, the corresponding complexes of thymidine and dihydroxypropoxyguanine (DHPG) with HSV1 and human TK were built. For the docking of the guanine derivative, the X-ray structure of Elongation Factor Tu (EF-Tu), co-crystallized with guanosine diphosphate, was taken as reference. Fitting of thymidine into the active sites was done with respect to similar interactions found in thymidylate kinase. To complement the analysis of the 3-D structures of the two kinases and the substrate enzyme interactions, site-directed mutagenesis of the thymidine recognition site of HSV1-TK has been undertaken, changing Asp162 in the thymidine recognition site into Asn. First investigations reveal that the enzymatic activity of the mutant protein is destroyed.

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Section: Resultsmentioning

confidence: 67%

“…-The stereochemical configuration of the open-chain sugar moiety (pro-chiral) has to be that of desoxy-ribose [44].…”

Section: Incorporation Of the Substrates Into The Active Sites Of Hsvmentioning

confidence: 99%

Computer-aided active-site-directed modeling of the Herpes Simplex Virus 1 and human thymidine kinase

Folkers

Trumpp-Kallmeyer

Gutbrod

et al. 1991

J Computer-Aided Mol Des

Self Cite

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Structural biology and diabetes mellitus: molecular pathogenesis and rational drug design

1992

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Emerging concepts in the aetiology and pathogenesis of Type 1 (insulin-dependent) diabetes mellitus may offer new opportunities for treatment and cure. Here we describe recent advances in structural molecular biology and molecular design relevant to rational drug discovery. Such approaches focus on the three-dimensional structures of macromolecules and their interactions. In the coming decade such techniques may be applied to a wide variety of diabetes-related targets.

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Herpesviral Deoxythymidine Kinases Contain a Site Analogous to the Phosphoryl-binding Arginine-rich Region of Porcine Adenylate Kinase; Comparison of Secondary Structure Predictions and Conservation

Balasubramaniam

Veerisetty

Gentry

1990

Journal of General Virology

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Twelve herpesviral deoxythymidine kinases were examined for regions of sequence similarity by multiple alignment. Six highly conserved sites were observed. Site 1 corresponded to a glycine-rich loop that forms part of the ATP-binding pocket in porcine adenylate kinase (PAK), and site 5 corresponded to a region in PAK, located on one lobe of the cleft, that contains arginine residues that bind substrate phosphoryl groups. Site 3, consisting of the motif-DRH-, is thought to be involved in thymine/deoxythymidine recognition; site 4, which is nearby, probably participates in this function as well. The functions of sites 2 and 6 have not been identified. Secondary structure predictions were made by the Garnier method and averaged for each position in the multiple alignment. The structure predicted for all six sites was typically a short flexible region (turn or coil) at or adjacent to the site, flanked by rigid structures (helix or sheet) on either side.

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Zur Entwicklung eines Pharmakophormodells für thymidinkinaseabhängige, nukleosidanaloge Virostatika

Cited by 5 publications

References 10 publications

Computer-aided active-site-directed modeling of the Herpes Simplex Virus 1 and human thymidine kinase

Computer-aided active-site-directed modeling of the Herpes Simplex Virus 1 and human thymidine kinase

Structural biology and diabetes mellitus: molecular pathogenesis and rational drug design

Herpesviral Deoxythymidine Kinases Contain a Site Analogous to the Phosphoryl-binding Arginine-rich Region of Porcine Adenylate Kinase; Comparison of Secondary Structure Predictions and Conservation

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