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Kinetic and structural characterization of an alternatively spliced variant of human mitochondrial 5'(3')-deoxyribonucleotidase
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SYSNO ASEP 0435125 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Kinetic and structural characterization of an alternatively spliced variant of human mitochondrial 5'(3')-deoxyribonucleotidase Author(s) Pachl, Petr (UMG-J)
Fábry, Milan (UMG-J) RID
Veverka, Václav (UOCHB-X) RID, ORCID
Brynda, Jiří (UMG-J) RID
Řezáčová, Pavlína (UMG-J) RIDSource Title Journal of Enzyme Inhibition and Medicinal Chemistry. - : Taylor & Francis - ISSN 1475-6366
Roč. 30, č. 1 (2015), 63-68Number of pages 6 s. Language eng - English Country GB - United Kingdom Keywords 5'(3')-deoxyribonucleotidase ; alternative splicing ; crystal structure ; hydrolase ; mitochondria Subject RIV EB - Genetics ; Molecular Biology R&D Projects GA203/09/0820 GA ČR - Czech Science Foundation (CSF) LK11205 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Institutional support UOCHB-X - RVO:61388963 ; UMG-J - RVO:68378050 UT WOS 000347956500010 DOI 10.3109/14756366.2013.879577 Annotation Abstract Human mitochondrial 5'(3')-deoxyribonucleotidase (mdN) catalyzes dephosphorylation of nucleoside monophosphates, and thus helps maintain homeostasis of deoxynucleosides required for mitochondrial DNA synthesis. Mature mdN is a 23-kDa dimeric protein with highest expression levels in the heart, brain and skeletal muscle. We have identified an alternative splice variant of the mdN gene containing an 18-nucleotide insertion encoding 6 amino acids (GKWPAT) at the 3'-end of the penultimate exon 4. We recombinantly expressed this enzyme variant and characterized its biochemical and kinetic properties as well as its three-dimensional structure. Our high-resolution (1.27 Å) crystal structure revealed that the insertion forms a loop located in the vicinity of the active site pocket and affects enzyme kinetic parameters as well as protein thermal stability. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2015
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