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Purification, crystallization and preliminary X-ray diffraction analysis of the kinase domain of human tousled-like kinase 2

Tousled-like kinases (TLKs) are an evolutionarily conserved family of serine/threonine protein kinases involved in chromatin dynamics, including DNA replication and repair, transcription and chromosome segregation. The two members of the family reported in humans, namely TLK1 and TLK2, localize to t...

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Autores principales: Garrote, Ana M., Redondo, Pilar, Montoya, Guillermo, Muñoz, Inés G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3944701/
https://www.ncbi.nlm.nih.gov/pubmed/24598926
http://dx.doi.org/10.1107/S2053230X14002581
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author Garrote, Ana M.
Redondo, Pilar
Montoya, Guillermo
Muñoz, Inés G.
author_facet Garrote, Ana M.
Redondo, Pilar
Montoya, Guillermo
Muñoz, Inés G.
author_sort Garrote, Ana M.
collection PubMed
description Tousled-like kinases (TLKs) are an evolutionarily conserved family of serine/threonine protein kinases involved in chromatin dynamics, including DNA replication and repair, transcription and chromosome segregation. The two members of the family reported in humans, namely TLK1 and TLK2, localize to the cell nucleus and are capable of forming homo- or hetero-oligomers by themselves. To characterize the role of TLK2, its C-terminal kinase domain was cloned and overexpressed in Escherichia coli followed by purification to homogeneity. Crystallization experiments in the presence of ATP-γ-S yielded crystals suitable for X-ray diffraction analysis belonging to two different space groups: tetragonal I4(1)22 and cubic P2(1)3. The latter produced the best diffracting crystal (3.4 Å resolution using synchrotron radiation), with unit-cell parameters a = b = c = 126.05 Å, α = β = γ = 90°. The asymmetric unit contained one protein molecule, with a Matthews coefficient of 4.59 Å(3) Da(−1) and a solvent content of 73.23%.
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spelling pubmed-39447012014-03-12 Purification, crystallization and preliminary X-ray diffraction analysis of the kinase domain of human tousled-like kinase 2 Garrote, Ana M. Redondo, Pilar Montoya, Guillermo Muñoz, Inés G. Acta Crystallogr F Struct Biol Commun Crystallization Communications Tousled-like kinases (TLKs) are an evolutionarily conserved family of serine/threonine protein kinases involved in chromatin dynamics, including DNA replication and repair, transcription and chromosome segregation. The two members of the family reported in humans, namely TLK1 and TLK2, localize to the cell nucleus and are capable of forming homo- or hetero-oligomers by themselves. To characterize the role of TLK2, its C-terminal kinase domain was cloned and overexpressed in Escherichia coli followed by purification to homogeneity. Crystallization experiments in the presence of ATP-γ-S yielded crystals suitable for X-ray diffraction analysis belonging to two different space groups: tetragonal I4(1)22 and cubic P2(1)3. The latter produced the best diffracting crystal (3.4 Å resolution using synchrotron radiation), with unit-cell parameters a = b = c = 126.05 Å, α = β = γ = 90°. The asymmetric unit contained one protein molecule, with a Matthews coefficient of 4.59 Å(3) Da(−1) and a solvent content of 73.23%. International Union of Crystallography 2014-02-19 /pmc/articles/PMC3944701/ /pubmed/24598926 http://dx.doi.org/10.1107/S2053230X14002581 Text en © Garrote et al. 2014 http://creativecommons.org/licenses/by/2.0/uk/ This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.
spellingShingle Crystallization Communications
Garrote, Ana M.
Redondo, Pilar
Montoya, Guillermo
Muñoz, Inés G.
Purification, crystallization and preliminary X-ray diffraction analysis of the kinase domain of human tousled-like kinase 2
title Purification, crystallization and preliminary X-ray diffraction analysis of the kinase domain of human tousled-like kinase 2
title_full Purification, crystallization and preliminary X-ray diffraction analysis of the kinase domain of human tousled-like kinase 2
title_fullStr Purification, crystallization and preliminary X-ray diffraction analysis of the kinase domain of human tousled-like kinase 2
title_full_unstemmed Purification, crystallization and preliminary X-ray diffraction analysis of the kinase domain of human tousled-like kinase 2
title_short Purification, crystallization and preliminary X-ray diffraction analysis of the kinase domain of human tousled-like kinase 2
title_sort purification, crystallization and preliminary x-ray diffraction analysis of the kinase domain of human tousled-like kinase 2
topic Crystallization Communications
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3944701/
https://www.ncbi.nlm.nih.gov/pubmed/24598926
http://dx.doi.org/10.1107/S2053230X14002581
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