Sortase A Assay Kit

• Background:

  Staphylococcal Sortase A is a bacterial transpeptidase that covalently attaches proteins to the bacterial cell wall, maintaining bacterial virulence and infectivity. Sortase A cleaves a specific peptide sequence (LPXTG recognition motif) within a target protein between threonine and glycine. The cysteine residue of the active site forms a transient thioacyl intermediate complex with the substrate protein. This intermediate complex and substrate is then immediately attacked by oligo-glycine nucleophiles present on peptide-glycans of the bacterial wall to form an amide bond.Since Sortase A is a critical enzyme needed to maintain the infectivity of the bacterium, it represents a promising therapeutic target. Especially for the treatment of infections caused by antibiotic-resistant bacteria.

• Description:

  The Sortase A Assay Kit is a fluorogenic assay kit designed to measure Sortase A proteolytic activity for screening and profiling applications.Figure 1: Illustration of the assay principle.The fluorescence emitted by donor DNP is quenched due to the proximity of the Abz acceptor in the intact peptide. The protease cleaves the peptide between T (threonine) and G (glycine) and forms an intermediate complex between the cysteine residue within the active site and the C-terminus of the peptide substrate.The cleaved substrateis then covalently boundedto the triglycine nucleophile. The glycine-DNP fragment is released, freeing the emission of fluorescence by DNP.Need us to run inhibitor screens or profile your compounds against Sortase A? Check out ourProtease Screening Services.

• UniProt:

  Q9S446

• Applications:

  Screen small molecules inhibitors of Sortase A in high-throughput applicationsTiter inhibitors of Sortase A

• Format:

  Catalog #NameAmountStorage71086Sortase A, His-Tag*12.5 μg-80°CAvoidmultiplefreeze/thawcycles!799382x Sortase assay buffer2.5 ml-20°C79939Triglycine250 μl-20°C79940Abz/Dnp substrate250 μl-20°C79685Low binding, black NUNC 96-well plate1Room Temp*The concentration of Sortase A is lot-specific and will be indicated on the tube containing the enzyme.

• Shipping Conditions:

  -80°C

• Storage Conditions:

  This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.