Open AccessDiazoacetyl Subtilisin
Author(s) -
Y. Stefanovsky,
F. H. Westheimer
Publication year - 1973
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.70.4.1132
Subject(s) - subtilisin , chemistry , hydrolysis , derivative (finance) , cleavage (geology) , enzyme , photodissociation , biochemistry , stereochemistry , organic chemistry , biology , paleontology , fracture (geology) , financial economics , economics
Subtilisin reacts at pH 6.8-7.8 withp -nitrophenyl diazoacetate to releasep -nitrophenol and form diazoacetyl subtilisin. Although at pH 7.8 this derivative rapidly undergoes hydrolytic cleavage to regenerate active enzyme, the derivative can be trapped by rapidly lowering the pH to 5. Similarly, with14 C-labeledp -nitrophenyl diazoacetate, the corresponding radiochemically labeled diazoacetyl enzyme can be prepared. Photolysis of this radioactive derivative incorporates radioactivity into the protein, and subsquent hydrolysis gives rise to several radioactive components, which, however, have not yet been identified.