a rat liver lysosome assay is used to monitor the extent of detoxification of a gliadin digest by caricain. Pre-incubating the gliadin digest for different durations with caricain allows the kinetics of the detoxification process to be studied. A significant degree of protection (80%) of the lysosomes is achieved with 1.7% w/w of caricain on substrate after incubation for 2 h at 37 °C. The detoxification follows first-order kinetics with a rate constant of 0.00017/sec
procaricain, fractions are estimated from plots of Kav against molecular weight using dextran blue, human gamma-globulin, bovine serum albumin and trypsin inhibitor as standards, mass spectrometric analysis of trypsin-digested fractions from chromatography are carried out by liquid chromatography/electrospray quadrupole time-of-flight mass spectrometry in positive mode
at low pH, enzyme undergoes conformational transition leading to instability and rapid degradation by pepsin. To be effective in gut after oral administration, enzyme needs to be protected against acid denaturation and degradation
Characterization of papaya peptidase A as a cysteine proteinase of Carica papaya L. with active-centre properties that differ from those of papain by using 2,2-dipyridyl disulphide and 4-chloro-7-nitrobenzofurazan as reactivity probes. Use of two-protonic-state electrophiles in the identification of catalytic-site thiol groups
Baines, B.S.; Brocklehurst, K.; Carey, P.R.; Jarvis, M.; Salih, E.; Storer, A.C.
Chymopapain A. Purification and investigation by covalent chromatography and characterization by two-protonic-state reactivity-probe kinetics, steady-state kinetics and resonance Raman spectroscopy of some dithioacyl derivatives
Subsite differences between the active centres of papaya peptidase A and papain as revealed by affinity chromatography. Purification of papaya peptidase A by ionic-strength-dependent affinity adsorption on an immobilized peptide inhibitor of papain
Topham, C.M.; Salih, E.; Frazao, C.; Kowlessur, D.; Overington, J.P.; Thomas, M.; Brocklehurst, S.M.; Patel, M.; Thomas, E.W.; Brocklehurst, K.
Structure-function relationships in the cysteine proteinases actinidin, papain and papaya proteinase omega. Three-dimensional structure of papaya proteinase omega deduced by knowledge-based modelling and active-centre characteristics determined by two-hydronic-state reactivity probe kinetics and kinetics of catalysis
Noble, M.A.; Gul, S.; Verma, C.S.; Brocklehurst, K.
Ionization characteristics and chemical influences of aspartic acid residue 158 of papain and caricain determined by structure-related kinetic and computational techniques: multiple electrostatic modulators of active-centre chemistry