A red-fluorescent substrate microarray for lipase fingerprinting.
A lipase substrate microarray was obtained by printing aliphatic C2-C12 monoesters of (5R)- and (5S)-3-(5,6-dihydroxyhexyloxy)benzaldehyde by reductive alkylation on amine-functionalized glass slides coated with bovine serum albumin and a short PEG linker. The microarray features 12 substrates and their 66 possible binary mixtures spotted in a 9 x 36 spot array. Lipase reactions are detected by chemoselective NaIO(4)-oxidation of the 1,2-diol hydrolysis product to form an aldehyde, which is then tagged with the red-fluorescent dye rhodamine B sulfohydrazide . Specific fingerprints are produced by active enzymes. These experiments provide the first example of lipase fingerprinting using microarrays.
Grognux
Johann
J
Department of Chemistry & Biochemistry, University of Berne, Freiestrasse 3, 3012 Berne, Switzerland.
Reymond
Jean-Louis
JL
D016000:Cluster Analysis; D005456:Fluorescent Dyes / Q000737:chemistry*; D008049:Lipase / Q000235:genetics*; D015394:Molecular Structure; D010449:Peptide Mapping / Q000379:methods*; D040081:Protein Array Analysis / Q000379:methods*; D013379:Substrate Specificity
D005456:Fluorescent Dyes; D008049:Lipase