[Serratia marcescens nucleases. I. Comparison of native and recombinant nucleases using electrospray mass spectrometry].
Isoforms of the natural and recombinant nucleases of Serratia marcescens were characterized by their molecular mass, which was determined by electrospray mass spectrometry. The natural nuclease was isolated from the S. marcescens B10M1 culture, whereas the recombinant nuclease was obtained from Escherichia coli MT102 cells carrying plasmid p403-SD2 with the nuclease gene nuc. The primary structure for each of the isoforms isolated from the nuclease preparations was determined by comparing its molecular mass with that of known amino acid sequence, which was determined from the nucleotide sequence of the nuc gene. Both preparations included identical nuclease variants with N-terminal amino acid residues removed. The number of isoforms in the natural nuclease was, however, significantly greater than in the recombinant nuclease. The structures of some of the isoforms were confirmed by N-terminal analysis.
Pedersen
J
J
Andersen
G
G
Roepstorff
P
P
Filimonova
M N
MN
Bidermann
K
K
D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D004706:Endodeoxyribonucleases / Q000737:chemistry* / Q000235:genetics; D004722:Endoribonucleases / Q000737:chemistry* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D007525:Isoelectric Focusing; D013058:Mass Spectrometry / Q000379:methods*; D008970:Molecular Weight; D010957:Plasmids; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics
D011994:Recombinant Proteins; D004706:Endodeoxyribonucleases; D004722:Endoribonucleases; C021835:Serratia marcescens nuclease
Nukleazy Serratia marcescens. I. Sravnenie prirodnoĭ i rekombinantnoĭ nukleaz s ispol'zovaniem élektrospreĭ-mass-spektrometrii.