Crystallization and data collection of the nucleotide-binding domain of Mg-ATPase.
Understanding of how P-type ATPases work would greatly benefit from the elucidation of more high-resolution structures. The nucleotide-binding domain of Mg-ATPase was selected for structural studies because Mg-ATPase is closely related to eukaryotic Ca-ATPase and Na,K-ATPase while the nucleotide-binding domain itself has diverged substantially. Two fragments of Mg-ATPase were cloned in Escherichia coli and purified. The entire cytoplasmic loop (residues 367-673), consisting of the phosphorylation and nucleotide-binding domains, expressed well and was purified in large quantities. The smaller 19.5 kDa nucleotide-binding domain (residues 383-545) expressed less well but formed crystals that diffracted to a resolution of 1.53 A which will be used for molecular replacement.
Håkansson
Kjell O
KO
Department of Biology, University of Copenhagen, Denmark. kohakansson@bio.ku.dk
Curović
Aida
A
D000251:Adenosine Triphosphatases / Q000737:chemistry*; D000595:Amino Acid Sequence; D017301:Ca(2+) Mg(2+)-ATPase / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry*; D026901:Membrane Transport Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D009711:Nucleotides / Q000378:metabolism*; D017434:Protein Structure, Tertiary
D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; D009711:Nucleotides; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase; C490561:MgtA protein, E coli