Increasing Stability and Toxicity of Pseudomonas Exotoxin by Attaching an Antiproteasic Peptide - CNRS - Centre national de la recherche scientifique Accéder directement au contenu
Article Dans Une Revue Biochemistry Année : 2011

Increasing Stability and Toxicity of Pseudomonas Exotoxin by Attaching an Antiproteasic Peptide

Résumé

Abbreviations used are: BPTI, bovine pancreatic trypsin inhibitor; DHFR, dihydrofolate reductase; DT, diphtheria, toxin; DTA, DT A-chain; DTB, DT B-chain; PE, Pseudomonas exotoxin A; RTA, ricin A-chain; rRTA, recombinant RTA; RTB, ricin B-chain. Abstract Trypsin-like activities are present within the endocytic pathway and allow cells to inactivate a fraction of incoming toxins, such as Pseudomonas exotoxin (PE), that require endocytic uptake before reaching the cytosol to inactivate protein synthesis. PE is a favorite toxin for building immunotoxins. The latter are promising molecules to fight cancer or transplant rejection, and producing more active toxins is a key challenge. More broadly, increasing protein stability is a potentially useful approach to improve the efficiency of therapeutic proteins. We report here that fusing an antiproteasic peptide (bovine pancreatic trypsin inhibitor, BPTI) to PE increases its toxicity to human cancer cell lines by 20-40 fold. Confocal microscopic examination of toxin endocytosis, digestion and immunoprecipitation experiments showed that the fused antiproteasic peptide specifically protects PE from trypsin-like activities. Hence, the attached BPTI acts as a bodyguard for the toxin within the endocytic pathway. Moreover, it increased the PE elimination half-time in mice by 70%, indicating that the fused BPTI stabilizes the toxin in vivo. This BPTI-fusion approach may be useful for protecting other circulating or internalized proteins of therapeutic interest from premature degradation.
Fichier principal
Vignette du fichier
Biochemistry2011.pdf (331.41 Ko) Télécharger le fichier
Origine : Fichiers produits par l'(les) auteur(s)

Dates et versions

hal-02137568 , version 1 (16-11-2020)

Identifiants

Citer

Jocelyn Méré, Christophe Chopard, Anne Bonhoure, Juliette Morlon-Guyot, Bruno Beaumelle. Increasing Stability and Toxicity of Pseudomonas Exotoxin by Attaching an Antiproteasic Peptide. Biochemistry, 2011, 50 (46), pp.10052-10060. ⟨10.1021/bi2010009⟩. ⟨hal-02137568⟩
27 Consultations
142 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More