Use of trinitrophenylation for quantification of protease and peptidase activities
Biochemistry (Mosc). 2001 May;66(5):531-4.
Abstract A sensitive and precise method for quantifying protease and peptidase activities is suggested.
N-Terminal amino groups of peptides which are formed during hydrolysis of the substrates react with
trinitrobenzenesulfonic acid (TNBS), and the trinitrophenyl (TNP) derivatives are determined spectrophotometrically.
Spontaneous hydrolysis of TNBS is considerably diminished on trinitrophenylation at pH 7.4 rather than at pH 9-10
as is usually used. The trinitrophenylation method can be used to determine the initial rate of hydrolysis and the
kinetics of reactions catalyzed by proteases and peptidases.
