title: Development of a Sortase A-mediated Peptide-labeled Liposome Applicable to Drug-delivery Systems.
abstract: OBJECTIVE
  In order to develop an efficient drug-delivery system (DDS), a lipopeptide-loaded liposome that functions as a platform for the transpeptidase reaction mediated by sortase A (SrtA) was constructed and its stability, as well as cell-specific targeting were evaluated in the present study.
  
  
  METHODS
  Several lipopeptides possessing an acceptor peptide sequence (oligoglycine ≥ three residues) or donor peptide sequence (LPETG) for the SrtA-mediated reaction were chemically synthesized and then inserted into the liposome membrane composed of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and cholesterol (DPPC-Chol-lipo) to obtain the lipopeptide-loaded liposomes. The transpeptidase reaction mediated by recombinant SrtA (His-ΔN59SrtA) was employed to modify the peptide moiety on the liposomal surface using a fluorescently-labeled substrate peptide corresponding to the species of each loaded lipopeptide. Furthermore, lung tumor-binding peptide (LTBP)-labeled liposomes, prepared by this transpeptidase reaction, were investigated for selective targeting to lung cancer cells in vitro.
  
  
  CONCLUSIONS
  The His-ΔN59SrtA-mediated transpeptidation of fluorescently-labeled peptide on the lipopeptide-loaded DPPC-Chol-lipo was confirmed. The selective targeting of LTBP-labeled liposomes to the lung cancer cell line A549 was also observed in vitro. These results suggest that the labeling of acceptor or donor lipopeptide-loaded liposomes with the transpeptidase SrtA could be a useful method for developing a platform applicable to a cancer-targeting DDS.
authors.affiliation: Department of Biological Science and Technology, Life System, Institute of Technology and Science, Tokushima University Graduate School, Tokushima, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Tabata
authors.affiliation: Department of Biological Science and Technology, Life System, Institute of Technology and Science, Tokushima University Graduate School, Tokushima, Japan.
authors.forename: Yukimasa
authors.initials: Y
authors.lastname: Ohkubo
authors.affiliation: Department of Biological Science and Technology, Life System, Institute of Technology and Science, Tokushima University Graduate School, Tokushima, Japan.
authors.forename: Natsuki
authors.initials: N
authors.lastname: Anyoji
authors.affiliation: Faculty of Pharmaceutical Sciences and Cooperative Research Center of Life Sciences, Kobe Gakuin University, Kobe, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Hojo
authors.affiliation: Department of Biological Science and Technology, Life System, Institute of Technology and Science, Tokushima University Graduate School, Tokushima, Japan.
authors.forename: Toshifumi
authors.initials: T
authors.lastname: Tomoyasu
authors.affiliation: Faculty of Pharmaceutical Sciences, Suzuka University of Medical Science, Suzuka, Mie, Japan.
authors.forename: Youhei
authors.initials: Y
authors.lastname: Tatematsu
authors.affiliation: Faculty of Pharmaceutical Sciences, Suzuka University of Medical Science, Suzuka, Mie, Japan.
authors.forename: Kazuto
authors.initials: K
authors.lastname: Ohkura
authors.affiliation: Department of Biological Science and Technology, Life System, Institute of Technology and Science, Tokushima University Graduate School, Tokushima, Japan nagamune@tokushima-u.ac.jp.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Nagamune
mesh_terms: D015060:1,2-Dipalmitoylphosphatidylcholine / Q000031:analogs & derivatives / Q000737:chemistry; D019881:Aminoacyltransferases / Q000737:chemistry*; D000970:Antineoplastic Agents / Q000008:administration & dosage*; D001426:Bacterial Proteins / Q000737:chemistry*; D045744:Cell Line, Tumor; D002784:Cholesterol / Q000737:chemistry / Q000378:metabolism; D003546:Cysteine Endopeptidases / Q000737:chemistry*; D016503:Drug Delivery Systems; D006801:Humans; D055666:Lipopeptides / Q000138:chemical synthesis / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry*; D008175:Lung Neoplasms / Q000188:drug therapy*
chemical_list: D000970:Antineoplastic Agents; D001426:Bacterial Proteins; D055666:Lipopeptides; D008081:Liposomes; D015060:1,2-Dipalmitoylphosphatidylcholine; C081581:colfosceril palmitate; D002784:Cholesterol; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases
keywords: Drug-delivery system; liposome; sortase A