Insight into catalytic mechanism of papain-like cysteine proteinases: the case of D158.
We studied the role of D158 in papain-like cysteine proteinases by using subtilisin Carlsberg, and its chemically modified analog thiolsubtilisin, by applying the proton inventory (PI) method and also by taking into account the pH profiles of the kcat/Km parameter. In the case of thiolsubtilisin, we estimated large inverse solvent isotope effects for kcat/Km, as in papain, whereas for subtilisin we found "dome-shaped" PI, suggesting a completely different mechanism. Finally, the kinetic behavior of thiolsubtilisin presented similarities as well as differences, compared to papain, suggesting a possible role for D158 as part of a catalytic triad in papain-like cysteine proteinases.
University of Ioannina, Department of Chemistry, 45110, Greece. epapamic@cc.uoi.gr
E M
EM
Papamichael
L G
LG
Theodorou
J G
JG
Bieth
D001224:Aspartic Acid / Q000378:metabolism*; D020134:Catalytic Domain; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D003627:Data Interpretation, Statistical; D007700:Kinetics; D010206:Papain / Q000378:metabolism; D020860:Subtilisin / Q000378:metabolism*
D001224:Aspartic Acid; D020860:Subtilisin; D003546:Cysteine Endopeptidases; D010206:Papain