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

Lisbeth Haue; Per A Pedersen; Peter L Jorgensen; Kjell O Håkansson

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

Lisbeth Haue, Per A Pedersen, Peter L Jorgensen, Kjell O Håkansson

6 Citations (Scopus)

Abstract

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.

Original language	English
Journal	Acta crystallographica Section D: Structural biology
Volume	59
Issue number	Pt 7
Pages (from-to)	1259-61
Number of pages	3
ISSN	0907-4449
Publication status	Published - Jul 2003

Keywords

Amino Acid Sequence
Animals
Binding Sites
Cloning, Molecular
Crystallization/methods
Membrane Proteins/chemistry
Nickel
Protein Structure, Tertiary
Protein Subunits/chemistry
Sodium-Potassium-Exchanging ATPase/chemistry
Swine
Thimerosal
X-Ray Diffraction/methods

Cite this

Cloning, expression, purification and crystallization of the N-domain from the alpha(2) subunit of the membrane-spanning Na,K-ATPase protein. / Haue, Lisbeth; Pedersen, Per A; Jorgensen, Peter L et al.
In: Acta crystallographica Section D: Structural biology , Vol. 59, No. Pt 7, 07.2003, p. 1259-61.

Research output: Contribution to journal › Journal article › Research › peer-review

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

abstract = "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.",

keywords = "Amino Acid Sequence, Animals, Binding Sites, Cloning, Molecular, Crystallization/methods, Membrane Proteins/chemistry, Nickel, Protein Structure, Tertiary, Protein Subunits/chemistry, Sodium-Potassium-Exchanging ATPase/chemistry, Swine, Thimerosal, X-Ray Diffraction/methods",

author = "Lisbeth Haue and Pedersen, {Per A} and Jorgensen, {Peter L} and H{\aa}kansson, {Kjell O}",

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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 - 0907-4449

VL - 59

SP - 1259

EP - 1261

JO - Acta crystallographica Section D: Structural biology

JF - Acta crystallographica Section D: Structural biology

IS - Pt 7

ER -

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

Abstract

Keywords

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