Purification and characterization of the deoxynucleoside monophosphate kinase of bacteriophage T5.
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.
Mikoulinskaia
Galina V
GV
Pushchino Branch of the Institute of Bioorganic Chemistry, Russian Academy of Sciences, ave Nauki 6, Pushchino, Moscow region 142290, Russia.
Gubanov
Sergei I
SI
Zimin
Andrei A
AA
Kolesnikov
Igor V
IV
Feofanov
Sergei A
SA
Miroshnikov
Anatolii I
AI
D000595:Amino Acid Sequence; D000645:Ammonium Sulfate / Q000494:pharmacology; D000837:Anion Exchange Resins / Q000494:pharmacology; D001435:Bacteriophages / Q000201:enzymology*; D002852:Chromatography, Ion Exchange; D003854:Deoxyribonucleotides / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D008956:Models, Chemical; D008969:Molecular Sequence Data; D017856:Phosphotransferases (Phosphate Group Acceptor) / Q000737:chemistry* / Q000302:isolation & purification*; D017434:Protein Structure, Tertiary; D012117:Resins, Synthetic; D012984:Software; D013997:Time Factors; D014461:Ultracentrifugation
D000837:Anion Exchange Resins; D003854:Deoxyribonucleotides; D012117:Resins, Synthetic; C042855:Mono Q; D017856:Phosphotransferases (Phosphate Group Acceptor); C020015:(deoxy)nucleoside-phosphate kinase; D000645:Ammonium Sulfate