Two isoforms of Serratia marcescens nuclease. Crystallization and preliminary X-ray investigation of the enzyme.
Two isoforms of an extracellular endonuclease, nucleases Sm1 and Sm2, were purified from culture fluid of Serratia marcescens strain BIO MI by ligand-exchange chromatography on phosphocellulose and DEAE-Toyopearl 650S. The pI-values for nucleases Sm1 and Sm2 were found to be 7.1 and 6.7, respectively. The amino acid analysis and N-terminal amino acid sequencing of the proteins showed a significant degree of homology between the enzymes. The nuclease Sm1 has been crystallized from ammonium sulfate solution by the vapour diffusion technique. The crystals belong to the space group P2(1)2(1)2(1) with unit cell constants a = 69.0, b = 106.7, c = 74.8 A, contain two molecules in an asymmetric unit, packing density Vm = 2.3 A/Da, and diffract to at least 1.5 A resolution. The Pt- and UO2-derivatives of the protein were obtained. Preliminary X-ray investigation of nuclease Sm2 crystals was carried out.
Bannikova
G E
GE
Bioengineriya, Centre, Institute of Molecular Biology, USSR Academy of Sciences, Moscow.
Blagova
E V
EV
Dementiev
A A
AA
Morgunova EYu
Mikchailov
A M
AM
Shlyapnikov
S V
SV
Varlamov
V P
VP
Vainshtein
B K
BK
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002852:Chromatography, Ion Exchange; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D004706:Endodeoxyribonucleases; D004720:Endonucleases / Q000737:chemistry*; D004722:Endoribonucleases; D007526:Isoelectric Point; D007527:Isoenzymes / Q000737:chemistry; D008969:Molecular Sequence Data; D012706:Serratia marcescens / Q000201:enzymology*; D014961:X-Ray Diffraction
D000596:Amino Acids; D007527:Isoenzymes; D004706:Endodeoxyribonucleases; D004720:Endonucleases; D004722:Endoribonucleases; C021835:Serratia marcescens nuclease