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Title: Expression, purification, crystallization and preliminary diffraction studies of the mammalian DAG kinase homologue YegS from Escherichia coli

Abstract

The overexpression, crystallization and preliminary diffraction analysis of E. coli YegS are reported. yegS is a gene encoding a 32 kDa cytosolic protein with unknown function but with strong sequence homology to a family of structurally uncharacterized eukaryotic non-protein kinases: diacylglycerol kinases, sphingosine kinases and ceramide kinases. Here, the overexpression, crystallization and preliminary diffraction analysis of Escherichia coli YegS are reported. The crystals belong to space group P2{sub 1}, with unit-cell parameters a = 42.4, b = 166.1, c = 48.5 Å, β = 96.97°. The presence of a dimer in the asymmetric unit was estimated to give a Matthews coefficient (V{sub M}) of 2.5 Å{sup 3} Da{sup −1} and a solvent content of 50.8%(v/v). Single-wavelength diffraction data were collected to a resolution of 1.9 Å using synchrotron radiation.

Authors:
 [1]; ;  [2];  [3]
  1. Department of Biochemistry and Biophysics, Stockholm University, SE-10691 Stockholm (Sweden)
  2. Department of Medical Biochemistry and Biophysics, Karolinska Institute, Scheeles väg 2, SE-171 77 Stockholm (Sweden)
  3. (Sweden)
Publication Date:
OSTI Identifier:
22356280
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 62; Journal Issue: Pt 3; Other Information: PMCID: PMC2197167; PMID: 16511327; PUBLISHER-ID: bw5128; OAI: oai:pubmedcentral.nih.gov:2197167; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; CRYSTALS; DIFFRACTION; DIMERS; ESCHERICHIA COLI; RESOLUTION; SOLVENTS; SPACE GROUPS; SYNCHROTRON RADIATION; WAVELENGTHS

Citation Formats

Bakali H, M. Amin, Nordlund, Pär, Hallberg, B. Martin, E-mail: martin.hallberg@mbb.ki.se, and Department of Biochemistry and Biophysics, Stockholm University, SE-10691 Stockholm. Expression, purification, crystallization and preliminary diffraction studies of the mammalian DAG kinase homologue YegS from Escherichia coli. United Kingdom: N. p., 2006. Web. doi:10.1107/S1744309106004799.
Bakali H, M. Amin, Nordlund, Pär, Hallberg, B. Martin, E-mail: martin.hallberg@mbb.ki.se, & Department of Biochemistry and Biophysics, Stockholm University, SE-10691 Stockholm. Expression, purification, crystallization and preliminary diffraction studies of the mammalian DAG kinase homologue YegS from Escherichia coli. United Kingdom. doi:10.1107/S1744309106004799.
Bakali H, M. Amin, Nordlund, Pär, Hallberg, B. Martin, E-mail: martin.hallberg@mbb.ki.se, and Department of Biochemistry and Biophysics, Stockholm University, SE-10691 Stockholm. Wed . "Expression, purification, crystallization and preliminary diffraction studies of the mammalian DAG kinase homologue YegS from Escherichia coli". United Kingdom. doi:10.1107/S1744309106004799.
@article{osti_22356280,
title = {Expression, purification, crystallization and preliminary diffraction studies of the mammalian DAG kinase homologue YegS from Escherichia coli},
author = {Bakali H, M. Amin and Nordlund, Pär and Hallberg, B. Martin, E-mail: martin.hallberg@mbb.ki.se and Department of Biochemistry and Biophysics, Stockholm University, SE-10691 Stockholm},
abstractNote = {The overexpression, crystallization and preliminary diffraction analysis of E. coli YegS are reported. yegS is a gene encoding a 32 kDa cytosolic protein with unknown function but with strong sequence homology to a family of structurally uncharacterized eukaryotic non-protein kinases: diacylglycerol kinases, sphingosine kinases and ceramide kinases. Here, the overexpression, crystallization and preliminary diffraction analysis of Escherichia coli YegS are reported. The crystals belong to space group P2{sub 1}, with unit-cell parameters a = 42.4, b = 166.1, c = 48.5 Å, β = 96.97°. The presence of a dimer in the asymmetric unit was estimated to give a Matthews coefficient (V{sub M}) of 2.5 Å{sup 3} Da{sup −1} and a solvent content of 50.8%(v/v). Single-wavelength diffraction data were collected to a resolution of 1.9 Å using synchrotron radiation.},
doi = {10.1107/S1744309106004799},
journal = {Acta Crystallographica. Section F},
number = Pt 3,
volume = 62,
place = {United Kingdom},
year = {Wed Mar 01 00:00:00 EST 2006},
month = {Wed Mar 01 00:00:00 EST 2006}
}
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