Purification and characterization of the deoxynucleoside monophosphate kinase of bacteriophage T5.
Protein Expr Purif, 2003/2;27(2):195-201.
Mikoulinskaia GV[1], Gubanov SI, Zimin AA, Kolesnikov IV, Feofanov SA, Miroshnikov AI
Affiliations
PMID: 12597877
Impact factor: 2.025
Abstract
Deoxynucleoside monophosphate kinase (dNMP kinase) of bacteriophage T5 (EC 2.7.4.13) was purified to apparent homogeneity from phage-infected Escherichia coli cells. Electrophoresis in sodium dodecyl sulfate-polyacrylamide gel showed that the enzyme has a molecular mass of about 29 kDa. The molecular mass of dNMP kinase estimated by analytical equilibrium ultracentrifugation turned out to be 29.14 +/- 3.03 kDa. These data suggest that the enzyme exists in solution as a monomer. The isoelectric point of dNMP kinase was found to be 4.2. The N-terminal amino acid sequence, comprising 21 amino acids, was determined to be VLVGLHGEAGSGKDGVAKLII. A comparison of this amino acid sequence and those of known enzymes with a similar function suggests the presence of a nucleotide-binding site in the sequenced region.
MeSH terms
Amino Acid Sequence; Ammonium Sulfate; Anion Exchange Resins; Bacteriophages; Chromatography, Ion Exchange; Deoxyribonucleotides; Electrophoresis, Polyacrylamide Gel; Escherichia coli; Hydrogen-Ion Concentration; Models, Chemical; Molecular Sequence Data; Phosphotransferases (Phosphate Group Acceptor); Protein Structure, Tertiary; Resins, Synthetic; Software; Time Factors; Ultracentrifugation
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