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Effect of valine 106 on structure-function relation of cytosolic human thymidine kinase - Kinetic properties and oligomerization pattern of nine substitution mutants of V106. / Frederiksen, Hanne; Berenstein, Dvora; Munch-Petersen, Birgitte.

In: European Journal of Biochemistry, Vol. 271, No. 11, 2004, p. 2248-2256.

Publication: Research - peer-reviewJournal article – Annual report year: 2004

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Frederiksen, Hanne; Berenstein, Dvora; Munch-Petersen, Birgitte / Effect of valine 106 on structure-function relation of cytosolic human thymidine kinase - Kinetic properties and oligomerization pattern of nine substitution mutants of V106.

In: European Journal of Biochemistry, Vol. 271, No. 11, 2004, p. 2248-2256.

Publication: Research - peer-reviewJournal article – Annual report year: 2004

Bibtex

@article{5916b568706645bba84c987901d9003d,
title = "Effect of valine 106 on structure-function relation of cytosolic human thymidine kinase - Kinetic properties and oligomerization pattern of nine substitution mutants of V106",
publisher = "Springer Verlag",
author = "Hanne Frederiksen and Dvora Berenstein and Birgitte Munch-Petersen",
year = "2004",
doi = "10.1111/j.1432-1033.2004.04166.x",
volume = "271",
number = "11",
pages = "2248--2256",
journal = "European Journal of Biochemistry",
issn = "0014-2956",

}

RIS

TY - JOUR

T1 - Effect of valine 106 on structure-function relation of cytosolic human thymidine kinase - Kinetic properties and oligomerization pattern of nine substitution mutants of V106

A1 - Frederiksen,Hanne

A1 - Berenstein,Dvora

A1 - Munch-Petersen,Birgitte

AU - Frederiksen,Hanne

AU - Berenstein,Dvora

AU - Munch-Petersen,Birgitte

PB - Springer Verlag

PY - 2004

Y1 - 2004

N2 - Information on the regulation and structure-function relation of enzymes involved in DNA precursor synthesis is pivotal, as defects in several of these enzymes have been found to cause depletion or deletion of mitochondrial DNA resulting in severe diseases. Here, the effect of amino acid 106 on the enzymatic properties of the cell-cycle-regulated human cytosolic thymidine kinase 1 (TK1) is investigated. On the basis of the previously observed profound differences between recombinant TK1 with Val106 (V106WT) and Met106 (V106M) in catalytic activity and oligomerization pattern, we designed and characterized nine mutants of amino acid 106 differing in size, conformation and polarity. According to their oligomerization pattern and thymidine kinetics, the TK1 mutants can be divided into two groups. Group I (V106A, V106I and V106T) behaves like V106WT, in that pre-assay exposure to ATP induces reversible transition from a dimer with low catalytic activity to a tetramer with high catalytic activity. Group II (V106G, V106H, V106K, V106L and V106Q) behaves like V106M in that they are permanently high activity tetramers, irrespective of ATP exposure. We conclude that size and conformation of amino acid 106 are more important than polarity for the catalytic activity and oligomerization of TK1. The role of amino acid 106 and the sequence surrounding it for dimer-tetramer transition was confirmed by cloning the putative interface fragment of human TK1 and investigating its oligomerization pattern.

AB - Information on the regulation and structure-function relation of enzymes involved in DNA precursor synthesis is pivotal, as defects in several of these enzymes have been found to cause depletion or deletion of mitochondrial DNA resulting in severe diseases. Here, the effect of amino acid 106 on the enzymatic properties of the cell-cycle-regulated human cytosolic thymidine kinase 1 (TK1) is investigated. On the basis of the previously observed profound differences between recombinant TK1 with Val106 (V106WT) and Met106 (V106M) in catalytic activity and oligomerization pattern, we designed and characterized nine mutants of amino acid 106 differing in size, conformation and polarity. According to their oligomerization pattern and thymidine kinetics, the TK1 mutants can be divided into two groups. Group I (V106A, V106I and V106T) behaves like V106WT, in that pre-assay exposure to ATP induces reversible transition from a dimer with low catalytic activity to a tetramer with high catalytic activity. Group II (V106G, V106H, V106K, V106L and V106Q) behaves like V106M in that they are permanently high activity tetramers, irrespective of ATP exposure. We conclude that size and conformation of amino acid 106 are more important than polarity for the catalytic activity and oligomerization of TK1. The role of amino acid 106 and the sequence surrounding it for dimer-tetramer transition was confirmed by cloning the putative interface fragment of human TK1 and investigating its oligomerization pattern.

U2 - 10.1111/j.1432-1033.2004.04166.x

DO - 10.1111/j.1432-1033.2004.04166.x

JO - European Journal of Biochemistry

JF - European Journal of Biochemistry

SN - 0014-2956

IS - 11

VL - 271

SP - 2248

EP - 2256

ER -