[Effect of thiol-specific reagents on the activity of PaeI and PaeII restrictases from Pseudomonas aeruginosa].
5,5'-dithiobis-2-nitrobenzoic acid, N-ethylmaleimide, and parachloromercuribenzoate have been demonstrated to inhibit the activity of restrictases PaeI and PaeII from Ps. aeruginosa bacterial cells. Restrictase PaeII was more sensitive to the action of thiol-specific reagents, as compared to PaeI. The minimal concentration of reagents for SH-groups that completely inhibited the activity of restrictases PaeI and PaeII was determined. The protective effect against the inhibitory action of 5,5'-dithiobis-2-nitrobenzoic acid on the activity of PaeII was observed after preincubation of these enzymes with phage lambda DNA and Mg2+ cations. It is suggested that restrictase PaeI and PaeII molecules contain SH-groups, essential for the enzymatic activity. They are believed responsible for restrictase binding with DNA substrate.
D010582:Bacteriophage lambda / Q000187:drug effects / Q000378:metabolism; D002850:Chromatography, Gel; D004262:DNA Restriction Enzymes / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D004279:DNA, Viral / Q000187:drug effects / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific; D004587:Electrophoresis, Agar Gel; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000201:enzymology; D013329:Structure-Activity Relationship; D013439:Sulfhydryl Reagents / Q000494:pharmacology*
D004279:DNA, Viral; D013439:Sulfhydryl Reagents; D004262:DNA Restriction Enzymes; C046184:endodeoxyribonuclease PaeI; C025651:CCCGGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific
Vliianie tiolspetsificheskikh reagnetov na aktivnost' restritaz PaeI i PaeII iz Pseudomonas aeruginosa.