Sortases are a family of membrane–anchored transpeptidases expressed by Gram–positive bacteria (1). Sortase A catalyzes the cleavage of C-terminal recognition motif (LPXTG) of multiple structurally unrelated proteins followed by formation of an amide bond with the peptidoglycan layer of the bacteria (2). Since sortases are widely distributed among a variety of bacterial pathogens and required for virulence, they represent a promising therapeutic target for the development of novel anti-infective agents (3). Sortase A has also been used as a biotechnology tool for a variety of protein modifications and immobilization by sortase-mediated protein ligation (4).
The SensoLyte® 520 Sortase A Assay Kit employs a novel internally quenched 5-FAM/QXL® FRET (Fluorescence Resonance Energy Transfer) substrate for the detection of protease activity. The enzyme cleaves the FRET substrate into two separate fragments resulting in the release of 5-FAM fluorescence, monitored at excitation/emission=
1. Spirig, T. et al. Mol Microbiol 82, 1044 (2011).
2. Marraffini, LA. et al. Microbiol Mol Biol Rev 70, 192 (2006).
3. Maresso, AW. et al. Pharmacol Rev 60, 128 (2008).
4. Proft, T. Biotechnol Lett 32, 1 (2010).
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AnaSpec, EGT Group
AnaSpec, EGT Group is a leading provider of integrated proteomics and genomics solutionsTM for worldwide life science research. AnaSpec offers expertise in peptides, detection reagents, antibodies, assay kits, amino acids, oligonucleotides, and qPCR MasterMixes. AnaSpec carries a broad product line of biochemicals and reagents for basic research, high-throughput screening and drug discovery. AnaSpec also provides premier custom services including peptide synthesis, antibody production, assay development, fluorescent dyes, and oligonucleotide synthesis. AnaSpec is certified to ISO 9001:2008.
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