Restriction endonuclease SsoII with photoregulated activity - A "molecular gate" approach

Le Thi Hien, Timofei S. Zatsepin, Benno Schierling, Eugene M. Volkov, Wolfgang Wende, Alfred Pingoud, Elena A. Kubareva, Tatiana S. Oretskaya

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1366-1373
Number of pages8
JournalBioconjugate Chemistry
Volume22
Issue number7
DOIs
Publication statusPublished - 20 Jul 2011
Externally publishedYes

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