title: Delivery of Non-Native Cargo into Mammalian Cells Using Anthrax Lethal Toxin.
abstract: The intracellular delivery of peptide and protein therapeutics is a major challenge due to the plasma membrane, which acts as a barrier between the extracellular environment and the intracellular milieu. Over the past two decades, a nontoxic PA/LFN delivery platform derived from anthrax lethal toxin has been developed for the transport of non-native cargo into the cytosol of cells in order to understand the translocation process through a protective antigen (PA) pore and to probe intracellular biological functions. Enzyme-mediated ligation using sortase A and native chemical ligation are two facile methods used to synthesize these non-native conjugates, inaccessible by recombinant technology. Cargo molecules that translocate efficiently include enzymes from protein toxins, antibody mimic proteins, and peptides of varying lengths and non-natural amino acid compositions. The PA pore has been found to effectively convey over 30 known cargos other than native lethal factor (LF; i.e., non-native) with diverse sequences and functionalities on the LFN transporter protein. All together these studies demonstrated that non-native cargos must adopt an unfolded or extended conformation and contain a suitable charge composition in order to efficiently pass through the PA pore. This review provides insight into design parameters for the efficient delivery of new cargos using PA and LFN.
authors.affiliation: Massachusetts Institute of Technology , Department of Chemistry, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.
authors.forename: Amy E
authors.initials: AE
authors.lastname: Rabideau
authors.affiliation: Massachusetts Institute of Technology , Department of Chemistry, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.
authors.forename: Bradley Lether
authors.initials: BL
authors.lastname: Pentelute
mesh_terms: D019881:Aminoacyltransferases / Q000737:chemistry; D000818:Animals; D000942:Antigens, Bacterial / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry; D001427:Bacterial Toxins / Q000737:chemistry*; D045744:Cell Line, Tumor; D003546:Cysteine Endopeptidases / Q000737:chemistry; D003600:Cytosol / Q000378:metabolism; D004337:Drug Carriers / Q000737:chemistry*; D006801:Humans; D026901:Membrane Transport Proteins / Q000378:metabolism; D021381:Protein Transport; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004337:Drug Carriers; D026901:Membrane Transport Proteins; D011506:Proteins; C030325:anthrax toxin; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases