Optimization of detergents for the assay of cathepsins B, L, S, and K.
The differential effects of representative, commonly available ionic (SDS), nonionic (Brij 35, Tween 20, and Triton X-100), and zwitterionic (Chaps) detergents on the catalytic activity and properties of human cathepsins B, L, S, and K were examined. The presence of detergents in the assay buffer affected the activity of cathepsins to variable extents; Chaps enhanced the activity of all the enzymes while SDS was most detrimental. Tween 20 lowered cathepsin S activity, while it slightly enhanced that of all other cathepsins studied. The presence of detergents in the activation buffer was clearly beneficial to both cathepsins L and K, possibly by favoring the release of the enzyme from the walls of the incubation vessel. Overall, the results indicate that Chaps is the optimal detergent for use with this family of enzymes.
Department of Surgery, McGill University, Montréal, Québec, Canada. jkrupa@shriners.mcgill.ca
J C
JC
Krupa
J S
JS
Mort
D002401:Cathepsin B / Q000032:analysis*; D056657:Cathepsin K; D056668:Cathepsin L; D002403:Cathepsins / Q000032:analysis*; D002793:Cholic Acids / Q000494:pharmacology; D003546:Cysteine Endopeptidases; D003902:Detergents / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D010450:Endopeptidases; D006801:Humans; D007700:Kinetics; D017830:Octoxynol / Q000494:pharmacology; D000077423:Polidocanol; D011092:Polyethylene Glycols / Q000494:pharmacology; D011136:Polysorbates / Q000494:pharmacology; D011994:Recombinant Proteins / Q000032:analysis; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013050:Spectrometry, Fluorescence
D002793:Cholic Acids; D003902:Detergents; D011136:Polysorbates; D011994:Recombinant Proteins; D000077423:Polidocanol; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D017830:Octoxynol; D002403:Cathepsins; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; C534281:CTSL protein, human; D056668:Cathepsin L; C028217:cathepsin S; C534130:CTSK protein, human; D056657:Cathepsin K; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate