Fractionation and purification of DNA methylases of Shigella sonnei

Journal Article · Sat May 10 04:00:00 EDT 1986 · Biochemistry (Engl. Transl.); (United States)

OSTI ID:5879086

Suchkov, S V; Nikol'skaya, I I; Debov, S S

A comparative study was made of the possibilities of various types of column chromatography and isoelectrofocusing for the analysis of the set of DNA methylases of the strains Shigella sonnei 47. On the basic of cation-exchange chromatography, a simple, rapid, and convenient method has been developed for the production of a highly active summary preparations of DNA methylases. It has been shown that affinity chromatography on heparin-Sepharose provides for the separation of methylases according to the characteristic of specificity for the nitrogen base. The presence of six individuals enzymes of DNA methylation, differing in value of the isoelectric point, was demonstrated in Shigella sonnei 47 cells by the IEF* method. Increased ability of the DNA methylases of Shigella sonnei 47 for nonspecific aggregation during the process of fractionation was demonstrated, which makes the use of column chromatography unsuitable for the isolation and purification of individual methylating enzymes of the investigated strain. The advantages of the IEF method, as well as its potentialities from the standpoint of studying various molecular forms of site-specific enzymes, are discussed.

• Research Organization: Scientific-Research Institute of Medical Enzymology, Moscow, USSR
• OSTI ID: 5879086
• Journal Information: Biochemistry (Engl. Transl.); (United States), Journal Name: Biochemistry (Engl. Transl.); (United States) Vol. 50:11; ISSN BIORA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKYL RADICALS
AMINO ACIDS
BACTERIA
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
CHROMATOGRAPHY
DNA
DRUGS
ELECTROPHORESIS
ENZYME ACTIVITY
ENZYMES
ESTERASES
FILTRATION
HYDROLASES
LABELLED COMPOUNDS
LABELLING
LIPOTROPIC FACTORS
METHIONINE
METHYL RADICALS
METHYLATION
MICROORGANISMS
NUCLEASES
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PHOSPHODIESTERASES
PURIFICATION
RADICALS
RADIOENZYMATIC ASSAY
SEPARATION PROCESSES
SHIGELLA
TRANSFERASES
TRITIUM COMPOUNDS