A hydrophilic azobenzene-bearing amino acid for photochemical control of a restriction enzyme BamHI.
A novel hydrophilic and negatively charged azobenzene-bearing amino acid, 4'-carboxyphenylazophenylalanine (azoAla 1), has been designed and synthesized for investigation of the photochemical regulation of the enzyme activity. The properties of photoisomerization and thermal stability of the cis-isomer were similar to those of a commonly used phenylazophenylalanine (azoAla 2). For photochemical control of the enzyme, these two azobenzene-bearing amino acids were incorporated into the specific position at the dimer interface of a restriction enzyme BamHI. These trans-azobenzene derivatives in the BamHI suppressed the enzymatic activity, and the following photoirradiation at 366 nm induced the recovery of its activity. Although the activities of both azoAla-BamHI mutants were same level after a long time irradiation, the recovery of the activity of azoAla 1-BamHI was faster than that of azoAla 2-BamHI with a short time irradiation. This result suggests that the negatively charged carboxylate group introduced into an azobenzene moiety affects the behavior of azoAla in the protein scaffold during the trans-cis photoisomerization.
Nakayama
Koji
K
The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047, Japan.
Endo
Masayuki
M
Majima
Tetsuro
T
D001391:Azo Compounds; D004262:DNA Restriction Enzymes / Q000528:radiation effects; D015245:Deoxyribonuclease BamHI / Q000528:radiation effects*; D010649:Phenylalanine / Q000031:analogs & derivatives*; D010777:Photochemistry / Q000379:methods*; D055672:Static Electricity; D013329:Structure-Activity Relationship
C087328:4-phenylazophenylalanine; D001391:Azo Compounds; D010649:Phenylalanine; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI; C009850:azobenzene