Identification of the EcoKI and EcoR124I Type I restriction--modification enzyme subunits by non-equilibrium pH gradient two-dimensional gel electrophoresis
Jazyk angličtina Země Spojené státy americké Médium print
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
12630312
DOI
10.1007/bf02818664
Knihovny.cz E-zdroje
- MeSH
- 2D gelová elektroforéza MeSH
- DNA restrikčně-modifikační enzymy chemie MeSH
- Escherichia coli enzymologie MeSH
- izoelektrický bod MeSH
- místně specifická DNA-methyltransferasa (adenin-specifická) chemie MeSH
- molekulová hmotnost MeSH
- restrikční endonukleasy typu I chemie MeSH
- restrikční enzymy chemie MeSH
- western blotting MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- DNA modification methylase EcoKI MeSH Prohlížeč
- DNA restrikčně-modifikační enzymy MeSH
- EcoR124 modification methylase type I MeSH Prohlížeč
- endodeoxyribonuclease EcoKI MeSH Prohlížeč
- endodeoxyribonuclease EcoR124I MeSH Prohlížeč
- místně specifická DNA-methyltransferasa (adenin-specifická) MeSH
- restrikční endonukleasy typu I MeSH
- restrikční enzymy MeSH
Effectively optimized and reproducible procedure for monitoring the composition of type I restriction-modification endonucleases EcoKI and EcoR124I by non-equilibrium pH gradient two-dimensional (2-D) gel electrophoresis is described. Three subunits of the enzyme complex, which widely differ from one another in their isoelectric points and molar mass, were identified in crude cell extracts of E. coli. For the first time all three subunits of both EcoKI and EcoR124I were detected as distinct spots on a single 2-D gel. A sensitive immunoblotting procedure was suggested suitable for routine use in determining the identity of individual subunits. Potential application of this method for detailed studies of regulation of the function and stoichiometry of the enzyme complexes is discussed.
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