Cloning, expression, purification and crystallization of the N-domain from the alpha(2) subunit of the membrane-spanning Na,K-ATPase protein

TY - JOUR

T1 - Cloning, expression, purification and crystallization of the N-domain from the alpha(2) subunit of the membrane-spanning Na,K-ATPase protein

AU - Haue, Lisbeth

AU - Pedersen, Per A

AU - Jorgensen, Peter L

AU - Håkansson, Kjell O

PY - 2003/7

Y1 - 2003/7

N2 - The nucleotide-binding domain of the Na,K-ATPase ion pump was expressed with a His tag in Escherichia coli and purified. The soluble 24 kDa derivative consists of 214 amino-acid residues and was crystallized in the presence of NiCl(2). The crystals belong to space group F23, with unit-cell parameters a = b = c = 147.5 A, and diffract to 3.1 A. Complete data sets could be collected from native and thimerosal-treated crystals frozen in 50% sucrose. Five mercury positions were found and initial SIR phases calculated.

AB - The nucleotide-binding domain of the Na,K-ATPase ion pump was expressed with a His tag in Escherichia coli and purified. The soluble 24 kDa derivative consists of 214 amino-acid residues and was crystallized in the presence of NiCl(2). The crystals belong to space group F23, with unit-cell parameters a = b = c = 147.5 A, and diffract to 3.1 A. Complete data sets could be collected from native and thimerosal-treated crystals frozen in 50% sucrose. Five mercury positions were found and initial SIR phases calculated.

KW - Amino Acid Sequence

KW - Animals

KW - Binding Sites

KW - Cloning, Molecular

KW - Crystallization/methods

KW - Membrane Proteins/chemistry

KW - Nickel

KW - Protein Structure, Tertiary

KW - Protein Subunits/chemistry

KW - Sodium-Potassium-Exchanging ATPase/chemistry

KW - Swine

KW - Thimerosal

KW - X-Ray Diffraction/methods

M3 - Journal article

C2 - 12832778

SN - 2059-7983

VL - 59

SP - 1259

EP - 1261

JO - Acta Crystallographica Section D: Biological Crystallography

JF - Acta Crystallographica Section D: Biological Crystallography

IS - Pt 7

ER -