Purification and properties of the MboII, a class-IIS restriction endonuclease.
After five purification steps a homogeneous preparation of endonuclease MboII was obtained, and several properties of the enzyme were determined. MboII is a monomer, with Mr under native and denaturing conditions being 47-49 x 10(3) Da. Endonuclease MboII is a basic protein (pI 8.3) which remains active when Mg2+ is replaced by Mn2+, Co2+, Ca2+, or Fe2+. MboII exhibits a star activity in the presence of some of the following reagents or ions: DMSO, glycerol, ethanol (and Co2+ or Mn2+ at pH 6). MboII does not bend DNA and is heat sensitive, losing activity after 15 min at 50 degrees C.
Sektas
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M
Department of Microbiology, University of Gdansk, Poland.
Kaczorowski
T
T
Podhajska
A J
AJ
D002413:Cations, Divalent; D002845:Chromatography; D004247:DNA / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004586:Electrophoresis; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D016955:Moraxella bovis / Q000201:enzymology*; D009690:Nucleic Acid Conformation; D013696:Temperature
D002413:Cations, Divalent; D004247:DNA; C024413:endodeoxyribonuclease MboII; D015252:Deoxyribonucleases, Type II Site-Specific