title: Restriction endonuclease SsoII with photoregulated activity--a "molecular gate" approach.
abstract: A novel method for regulating the activity of homodimeric proteins--"molecular gate" approach--was proposed and its usefulness illustrated for the type II restriction endonuclease SsoII (R.SsoII) as a model. The "molecular gate" approach is based on the modification of R.SsoII with azobenzene derivatives, which allows regulating DNA binding and cleavage via illumination with light. R.SsoII variants with single cysteine residues introduced at selected positions were obtained and modified with maleimidoazobenzene derivatives. A twofold change in the enzymatic activity after illumination with light of wavelengths of 365 and 470 nm, respectively, was demonstrated when one or two molecules of azobenzene derivatives were attached to the R.SsoII at the entrance of or within the DNA-binding site.
authors.affiliation: Department of Chemistry, Lomonosov Moscow State University, Leninskie gory, Moscow 119991, Russia.
authors.forename: Le Thi
authors.initials: le T
authors.lastname: Hien
authors.forename: Timofei S
authors.initials: TS
authors.lastname: Zatsepin
authors.forename: Benno
authors.initials: B
authors.lastname: Schierling
authors.forename: Eugene M
authors.initials: EM
authors.lastname: Volkov
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Wende
authors.forename: Alfred
authors.initials: A
authors.lastname: Pingoud
authors.forename: Elena A
authors.initials: EA
authors.lastname: Kubareva
authors.forename: Tatiana S
authors.initials: TS
authors.lastname: Oretskaya
mesh_terms: D001391:Azo Compounds / Q000737:chemistry*; D001665:Binding Sites; D004247:DNA / Q000378:metabolism*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics; D008027:Light; D008301:Maleimides / Q000737:chemistry; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D001391:Azo Compounds; D008301:Maleimides; D050505:Mutant Proteins; C043592:maleimide; D004247:DNA; C045297:CCNGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific; C009850:azobenzene