title: Total chemical synthesis of the enzyme sortase A(ΔN59) with full catalytic activity.
abstract: The enzyme sortase A is a ligase which catalyzes transpeptidation reactions.1, 2 Surface proteins, including virulence factors, that have a C terminal recognition sequence are attached to Gly5 on the peptidoglycan of bacterial cell walls by sortase A.1 The enzyme is an important anti-virulence and anti-infective drug target for resistant strains of Gram-positive bacteria.2 In addition, because sortase A enables the splicing of polypeptide chains, the transpeptidation reaction catalyzed by sortase A is a potentially valuable tool for protein science.3 Here we describe the total chemical synthesis of enzymatically active sortase A. The target 148 residue polypeptide chain of sortase AΔN59 was synthesized by the convergent chemical ligation of four unprotected synthetic peptide segments. The folded protein molecule was isolated by size-exclusion chromatography and had full enzymatic activity in a transpeptidation assay. Total synthesis of sortase A will enable more sophisticated engineering of this important enzyme molecule.
authors.affiliation: Department of Chemistry, The University of Chicago, 929 East 57th Street, Chicago, IL 60637 (USA).
authors.forename: Fang-Kun
authors.initials: FK
authors.lastname: Deng
authors.forename: Liang
authors.initials: L
authors.lastname: Zhang
authors.forename: Ya-Ting
authors.initials: YT
authors.lastname: Wang
authors.forename: Olaf
authors.initials: O
authors.lastname: Schneewind
authors.forename: Stephen B H
authors.initials: SB
authors.lastname: Kent
mesh_terms: D000595:Amino Acid Sequence; D019881:Aminoacyltransferases / Q000138:chemical synthesis* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000138:chemical synthesis* / Q000378:metabolism*; D002384:Catalysis; D002850:Chromatography, Gel; D003546:Cysteine Endopeptidases / Q000138:chemical synthesis* / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism*; D017510:Protein Folding; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D001426:Bacterial Proteins; D010446:Peptide Fragments; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases
keywords: chemical ligation; chemical protein synthesis; convergent synthesis; enzyme catalysis; sortase A