title: Evidence for a two-state transition in papain that may have no close analogue in ficin. Differences in the disposition of cationic sites and hydrophobic binding areas in the active centres of papain and ficin.
abstract: The kinetics of the reactions of the active-centre thiol groups of papain (EC 3.4.22.2) and ficin (EC 3.4.22.3) with the two-protonic-state reactivity probes 2,2'-dipyridyl disulphide, n-propyl 2-pyridyl disulphide and 4-(N-aminoethyl 2'-pyridyl disulphide)- 7-nitrobenzo-2-oxa-1,3-diazole (compound I) were studied over a wide range of pH. Differences between the reactivities of ficin and papain towards the cationic forms of the alkyl 2-pyridyl disulphide probes suggest that ficin contains a cationic site without exact analogue in papain, and the striking difference in the shapes of the pH-rate profiles for the reactions of the two enzymes with compound (1) suggests differences in the mobilities or dispositions of the active-centre histidine imidazole groups with respect to relevant hydrophobic binding areas. The evidence from reactivity-probe studies that the papain catalytic mechanism involves substantial repositioning of the active-centre imidazole group during the catalytic act does not apply also to ficin. If ficin contains an aspartic acid residue analogous to aspartic acid-158 in papain, the pKa of its carboxy group is probably significantly lower than the pKa of the analogous group in papain.
mesh_terms: D015082:2,2'-Dipyridyl; D001583:Benzoxazoles; D001665:Binding Sites; D055598:Chemical Phenomena; D002621:Chemistry; D004220:Disulfides; D010450:Endopeptidases; D005361:Ficain; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008956:Models, Chemical; D010206:Papain; D011487:Protein Conformation; D011725:Pyridines
chemical_list: D001583:Benzoxazoles; D004220:Disulfides; D011725:Pyridines; D015082:2,2'-Dipyridyl; C011643:1,4-(N-2-aminoethyl-2'-pyridyl disulfide)-7-nitrobenzo-2-oxa-1,3-diazole; C016698:n-propyl 2-pyridyl disulfide; D010450:Endopeptidases; D010206:Papain; D005361:Ficain