Sortase A, Heptamutant Protein (tag-free)

Description

Tag-free Recombinant Sortase A, Heptamutant Protein – for site-specific protein labeling, antibody modification, and cell surface – conjugations

• Recombinant Staphylococcus aureus Sortase A, Heptamutant
• Highly purified and extremely active calcium independent transpeptidase
• Highest cost-efficiency
• Specificity: Site-specific linkage of proteins and antibodies with LPXTG motif to other proteins and molecules with glycine or aminomethylene motif
• Versatility: Ligation reaction is compatible with many types of small or large molecules (e.g. nanoparticles, tags such as fluorescent dyes or biotin, peptides, proteins, drugs and compounds, and even surfaces)
• Tag-free
  Staphylococcus aureus Sortase A, Heptamutant is a calcium-independent and highly active Sortase A variant with seven mutations (P94R, D160N, D165A, Y187L, E189R, K190E, K196T) and is the most universally applicable sortase derivative known. TriAltus Bioscience´s recombinant Sortase A, Heptamutant, is the enzyme preparation with the highest purity and activity as well as cost-efficiency commercially available.

This engineered enzyme enables site-specific protein modification and ligation (sortase-mediated tagging, sortagging) with exceptional precision and efficiency under mild and pysiological reaction conditions, which makes it applicable in vitro and in vivo. This transpeptidase cleaves between threonine and glycine in an LPXTG motif, enabling precise, site-specific linkage of proteins genetically engineered with a C-terminal LPXTG motif (which ensures correct protein folding and functionality) to arbitrary molecules with an oligoglycine or aminomethylene motif, which turns it into an ubiquitous research tool for protein modification. The enzymatic approach overcomes the limitations of conventional chemical conjugation methods, which are much harder to control and often compromise protein or antibody functionality.

The heptamutant variant of Sortase A is calcium-independent, making it effective in both intracellular and extracellular environments. Applications include site-specific protein labeling with molecules such as fluorescent dyes, biotin, and nucleic acids, protein-protein or protein-peptide fusions, peptide chimeras, antibody modification (e.g. antibody-drug conjugates/ADCs), protein cyclization, protein immobilization by anchoring to nanoparticles or surfaces, and live-cell surface conjugation.

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 a C-terminal LPXTG 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.

SPECIFICATIONS

PURITY | ~99% (SDS-PAGE gel analysis)

ENZYME SOURCE | Staphylococcus aureus Sortase A, Heptamutant is expressed in E. coli with no
residual amino acids on either the N or C terminus and is purified with CL7 / CLīM technology. The protein is provided tag-free.

QUANTITY | 500 ug

STORAGE/SHIPPING CONCENTRATION | 5 mg/mL

STORAGE BUFFER | 17.4 mM Tris HCl pH 8.0, 0.43 M NaCl, 20% glycerol

RECOMMENDED STORAGE CONDITIONS | -80°C

EXPIRATION | 6 months from receipt when stored as directed. Avoid repeated freeze/thaw cycles.