title: Ultrafiltration, a useful method for isolation of intermediates in native chemical ligation exemplified with the total synthesis of Sortase AΔN59.
abstract: In this paper, ultrafiltration was employed to facilitate the isolation of intermediates in native chemical ligation. Depending on the molecular weight cutoff of the membrane used, molecules with different sizes could be purified, separated, or concentrated by the ultrafiltration process. Total chemical synthesis of the polypeptide chain of the enzyme Sortase AΔN59 was used as an example of the application of ultrafiltration in chemical protein synthesis. Sortase A is a ligase that catalyzes transpeptidation reactions between proteins that have C-terminal LPXTG recognition sequence and Gly5- on the peptidoglycan of bacterial cell walls. Ultrafiltration technique facilitated synthesis of Sortase AΔN59 and was a promising tool in isolation of intermediates in native chemical ligation.
authors.affiliation: Department of Chemistry, The University of Chicago, 929 East 57th street, Chicago, IL, 60615, USA; Department of Biochemistry, School of Medicine Room1017, Jinan University, Huangpu West Road 601, Tianhe District, Guangzhou, Guangdong Province, 510000, China.
authors.forename: Fang-kun
authors.initials: FK
authors.lastname: Deng
mesh_terms: D019881:Aminoacyltransferases / Q000138:chemical synthesis*; D001426:Bacterial Proteins / Q000138:chemical synthesis*; D003546:Cysteine Endopeptidases / Q000138:chemical synthesis*; D010455:Peptides / Q000138:chemical synthesis; D014462:Ultrafiltration / Q000379:methods
chemical_list: D001426:Bacterial Proteins; D010455:Peptides; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases
keywords: Sortase A; chemical protein synthesis; native chemical ligation (NCL); ultrafiltration