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Structural and functional studies of enzymes in nucleotide metabolism

a detailed investigation of two enzymes and interaction profiling of FDA-approved nucleoside analog drugs with 23 enzymes

Egeblad, Louise (2011). Structural and functional studies of enzymes in nucleotide metabolism. Diss. (sammanfattning/summary) Uppsala : Sveriges lantbruksuniv., ; 2011:11
ISBN 978-91-576-7546-0
[Doctoral thesis]

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Abstract

Enzymes in nucleotide metabolism serve as the producers of the building blocks for DNA and RNA. From a medical perspective, nucleotide metabolism, and in particular salvage pathway enzymes, have attracted special interest, as nucleoside prodrugs given in the treatment of cancer and HIV are converted into their active metabolite forms by these enzymes.

In this thesis, two enzymes; uridine monophosphate kinase (UMPK) from Ureaplasma parvum (Up) and human phosphoribosyltransferase domain containing protein 1 (PRTFDC1), have been investigated. Furthermore, a nucleoside analog library (NAL) consisting of 45 FDA-approved nucleoside analogs has been developed.

The structure of Up-UMPK revealed that it was a hexamer. Kinetic constants were determined for UMP and ATP. UTP was a competitive inhibitor of UMP, and a non-competitive inhibitor of ATP. In contrast to other bacterial UMPKs, Up-UMPK was not activated by GTP.

PRTFDC1 is a homolog of hypoxanthine-guanine phosphoribosyltransferase (HPRT). Mutations in HPRT are associated with Lesch-Nyhan syndrome. The three-dimensional structures of PRTFDC1 and HRPT are very similar. Even though PRTFDC1 recognizes guanine and hypoxanthine as substrates, the functional turnover rates are less than 1% of the activity of HPRT.

NAL was screened using the high-throughput method, differential static light scattering (DSLS). An interaction profile of 23 enzymes involved in nucleotide metabolism and NAL was revealed. Interactions were detected for uridine phosphorylase 1 (UPP1) and guanine deaminase (GDA) with eight and six nucleoside prodrugs, respectively. The knowledge gained from this study can be important in the future search for drug lead candidates for UPP1 and GDA.

Authors/Creators:Egeblad, Louise
Title:Structural and functional studies of enzymes in nucleotide metabolism
Subtitle:a detailed investigation of two enzymes and interaction profiling of FDA-approved nucleoside analog drugs with 23 enzymes
Year of publishing :2011
Volume:2011:11
Number of Pages:81
Papers/manuscripts:
NumberReferences
I.Egeblad-Welin, L., Welin, M., Wang, L., Eriksson, S. (2007). Structural and functional investigations of Ureaplasma parvum UMP kinase – a potential antibacterial drug target. FEBS J 274, 6403-6414.
II.*Welin, M., *Egeblad, L., Johansson, A., Stenmark, P., Wang, L., Flodin, S., Nyman, T., Trésaugues, L., Kotenyova, T., Johansson, I., Eriksson, S., Eklund, H., Nordlund, P. (2010). Structural and functional studies of the human phosphoribosyltransferase domain containing protein 1. FEBS J 277, 4920-4930. * Shared first authorship
III.Egeblad, L., Welin, M., Johansson, A., Flodin, S., Gräslund, S., Wang, L., Eriksson, S., Nordlund, P. Interaction profiling of FDA-approved nucleoside analog drugs with 23 enzymes of human nucleotide metabolism. Manuscript.
Place of Publication:Uppsala
Publisher:Institutionen for anatomi, fysiologi och biokemi, Sveriges lantbruksuniversitet
ISBN for printed version:978-91-576-7546-0
Language:English
Publication Type:Doctoral thesis
Full Text Status:Public
Agris subject categories.:L Animal production > L50 Animal physiology and biochemistry
Subjects:(A) Swedish standard research categories 2011 > 1 Natural sciences > 106 Biological Sciences (Medical to be 3 and Agricultural to be 4) > Biochemistry and Molecular Biology
(A) Swedish standard research categories 2011 > 1 Natural sciences > 106 Biological Sciences (Medical to be 3 and Agricultural to be 4) > Biophysics
(A) Swedish standard research categories 2011 > 3 Medical and Health Sciences > 304 Medical Biotechnology > Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Agrovoc terms:metabolism, nucleotides, ureaplasma, phosphoribosyltransferase
Keywords:Nucleotide metabolism, Nucleoside analogs, Ureaplasma parvum, Uridine monophosphate kinase, Phosphoribosyltransferase domain containing protein 1, Nucleoside analog library, Differential static light scattering, Enzyme kinetics, Crystal structure
URN:NBN:urn:nbn:se:slu:epsilon-e-88
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-e-88
ID Code:8107
Department:(VH) > Dept. of Anatomy, Physiology and Biochemistry
Deposited By: Louise Egeblad
Deposited On:09 May 2011 11:03
Metadata Last Modified:02 Dec 2014 10:46

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