Enzyme fingerprints of activity, and stereo- and enantioselectivity from fluorogenic and chromogenic substrate arrays.
A series of stereochemically and structurally diverse fluorogenic and chromogenic substrates for hydrolytic enzymes has been synthesized and used to characterize enzyme activity profiles of esterases, lipases, proteases, peptidases, phosphatases, and epoxide hydrolases. The substrates used are particularly resilient to nonspecific reactions due to their mechanism of activation. The activities recorded with the individual substrates are therefore remarkably reproducible, and enable us to use the overall pattern of activity as a specific fingerprint for the enzyme sample. Fingerprints of activity, and enantio- and stereoselectivity are displayed as arrays of color-scale squares that are easily analyzed visually. Such fingerprints might be useful for quality control, enzyme discovery, and possibly for addressing the issue of functional convergence in enzymes.
Department of Chemistry and Biochemistry, University of Bern, Switzerland.
Denis
D
Wahler
Fabrizio
F
Badalassi
Paolo
P
Crotti
Jean-Louis
JL
Reymond
D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis / Q000378:metabolism; D002863:Chromogenic Compounds; D005456:Fluorescent Dyes; D005656:Fungal Proteins / Q000032:analysis / Q000378:metabolism; D006867:Hydrolases / Q000032:analysis* / Q000378:metabolism; D007700:Kinetics; D015336:Molecular Probe Techniques / Q000592:standards; D015394:Molecular Structure; D012680:Sensitivity and Specificity; D013237:Stereoisomerism
D001426:Bacterial Proteins; D002863:Chromogenic Compounds; D005456:Fluorescent Dyes; D005656:Fungal Proteins; D006867:Hydrolases