title: Proteolytic fingerprinting of complex biological samples using combinatorial libraries of fluorogenic probes.
abstract: Proteases have garnered interest as candidate biomarkers and therapeutic targets for many human diseases. A key challenge is the identification and characterization of disease-relevant proteases in the complex milieu of biological fluids such as serum, plasma, and bronchoalveolar lavage, in which a multitude of hydrolases act in concert. This unit describes a protocol to map the global proteolytic substrate specificities of complex biological samples using a concise combinatorial library of internally quenched fluorogenic peptide probes (IQFPs). This substrate profiling approach provides a global and quantitative comparison of protease specificities between different biological samples. Such a comparative analysis can lead to the identification of disease-specific 'fingerprints' of proteolytic activities with potential utility in diagnosis and therapy.
authors.affiliation: Center for Advanced Drug Research, Biosciences Division, SRI International, Harrisonburg, Virginia.
authors.forename: Kalyani
authors.initials: K
authors.lastname: Jambunathan
authors.affiliation: Center for Advanced Drug Research, Biosciences Division, SRI International, Harrisonburg, Virginia.
authors.forename: Douglas S
authors.initials: DS
authors.lastname: Watson
authors.affiliation: Center for Advanced Drug Research, Biosciences Division, SRI International, Harrisonburg, Virginia.
authors.forename: Krishna
authors.initials: K
authors.lastname: Kodukula
authors.affiliation: Center for Advanced Drug Research, Biosciences Division, SRI International, Harrisonburg, Virginia.
authors.forename: Amit K
authors.initials: AK
authors.lastname: Galande
mesh_terms: D000595:Amino Acid Sequence; D005456:Fluorescent Dyes / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000097:blood / Q000378:metabolism* / Q000652:urine; D019151:Peptide Library; D010455:Peptides / Q000032:analysis / Q000378:metabolism*; D059748:Proteolysis; D040901:Proteomics / Q000379:methods*; D013050:Spectrometry, Fluorescence / Q000379:methods; D013379:Substrate Specificity
chemical_list: D005456:Fluorescent Dyes; D019151:Peptide Library; D010455:Peptides; D010447:Peptide Hydrolases