Synthesis and Hydrolysis by Cysteine and Serine Proteases of Short Internally Quenched Fluorogenic Peptides

Positional-scanning combinatorial libraries of fluorescence resonance energy transfer peptides to define substrate specificity of carboxydipeptidases: assays with human cathepsin B

“…The greater preference of the S 0 1 subsite of cathepsin B for hydrophobic residues, with either aliphatic or aromatic structure, is very clear. These data are in accordance with earlier observations with substrates that permit endopeptidase activity [48] or carboxydipeptidase activity [28]. The complete resistance of the peptides containing Pro in this sublibrary is noteworthy due to the imide nature of its peptide bond with the P 1 amino acid.…”

Cysteine Cathepsins in Human Dentin-Pulp Complex

“…Enamel was removed with high-speed bur, and dentin was powdered by using a carbide bur irrigated with Tris-HCl 50 mmol/L, NaCl 100 mmol/L buffer, pH 7.4. Dentin powder was collected, homogenized with a Potter-Elvehjem tissue grinder in 1.0 mL of 50 mmol/L Tris-HCl buffer, pH 7.4, containing 0.1 mol/L NaCl at 4 C. Large debris was removed by centrifugation at 10,000g, 15 minutes at 4 C. The total cysteine cathepsin activities in the supernatant of the samples were monitored spectrofluorometrically by using the cysteine cathepsin-specific fluorogenic substrate Z-FR-MCA (Sigma) in a Hitachi F-2000 spectrofluorometer (Hitachi Scientific Instruments, Inc, Hialeah, FL), with the excitation and emission wavelengths of 380 and 460 nm, respectively (48). Before the assay, the cysteine proteinases were activated by incubation for 5 minutes at 25 C in 50 mmol/L sodium phosphate (pH 6.3), 200 mmol/L NaCl, 1 mmol/L ethylenediaminetetraacetic acid (EDTA), and 2 mmol/L dithiothreitol.…”

Progress in Spectroscopic Probes with Cleavable Active Bonds

“…Other donor-acceptor pairs used for designing FRET probes include tryptophan/dansyl [67][68][69][70], tryptophan/2,4-dinitrophenyl (Dnp) [71][72][73][74][75], 7-methoxycoumarin (Mca)/Dnp [76], o -aminobenzoyl/Dnp [77,78], o -aminobenzoyl/3-nitrotyrosine [79][80][81], and so on [82]. By using a fluorophore-quencher pair of Cy3 and Cy5, George and coworkers [42] developed a long-wavelength fluorescent probe to study the activity of matrix-metalloproteinase 3.…”