Staphylococcus aureus Sortase A (GenBank Accession No. AF162687), a.a. 26 - 206 (end) with N-terminal His-tag, MW = 21.7 kDa, expressed in an E. coli expression system.
Background
Staphylococcal Sortase A is a bacterial transpeptidase that covalently attaches proteins to the bacterial cell wall by cleaving between threonine and glycine at an LPXTG recognition motif and catalyzes the formation of an amide bond between the carboxyl-group of threonine and the amino-group of the cell-wall peptidoglycan. This chemistry has been exploited as a molecular "stapler" to site-specifically link proteins with the C-terminal LPETGX motif to other proteins or molecules possessing a glycine or aminomethylene motif. This reaction is applicable to proteins in solution and on the living cell surface.
Immunogen Region
26-206 (end)
Tag
N-terminal His-tag
Formulation
40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 8 mM imidazole, 0.04% Tween-20, and 20% glycerol
Species
Staphylococcus aureus
Application
Suitable for use with in vitro protein ligation reactions.
Notes
Assay was done in the reaction buffer (50 ul) containing 50 mM HEPES (pH=7.4), 150 mM NaCl, 5 mM CaCl2, 5 mM (Gly)3, 25 mM Abz/Dnp substrate, and Sortase A for 30 min at 30°C. Fluorescence intensity was measured at Ex320nm/Em420nm.
Avoid freeze/thaw cycles.
GenBank
AF162687
Mw(kda)
27.1 kDa
Synonyms
SrtA, sortase A
Uniprot
Q9S446
Shipping Temperature
-80°C (dry ice)
Format
Aqueous buffer solution
Storage
At least 6 months at -80°C.
Reference
1. Mazmanian, S.K., et al., Science. 1999 Jul 30,285(5428):760-3. 2. Kappel, K., et al., Protein Sci. 2012 Dec,21(12):1858-71.