Cloning, expression, purification and crystallization of the N-domain from the alpha(2) subunit of the membrane-spanning Na,K-ATPase protein.
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.
Haue
Lisbeth
L
Biomembrane Center, August Krogh Institute, Copenhagen University, Universitetsparken 13, 2100 Copenhagen OE, Denmark.
Pedersen
Per A
PA
Jorgensen
Peter L
PL
Håkansson
Kjell O
KO
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D003001:Cloning, Molecular; D003460:Crystallization / Q000379:methods; D008565:Membrane Proteins / Q000737:chemistry; D009532:Nickel; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000737:chemistry; D000254:Sodium-Potassium-Exchanging ATPase / Q000737:chemistry*; D013552:Swine; D013849:Thimerosal; D014961:X-Ray Diffraction / Q000379:methods
D008565:Membrane Proteins; D021122:Protein Subunits; D013849:Thimerosal; C022838:nickel chloride; D009532:Nickel; D000254:Sodium-Potassium-Exchanging ATPase