Journal article

Selective purification of recombinant neuroactive peptides using the flagellar type III secretion system

  • Singer, Hanna M. Département de Médecine, Université de Fribourg, Switzerland
  • Erhardt, Marc Département de Médecine, Université de Fribourg, Switzerland
  • Steiner, Andrew M. Department of Medicinal Chemistry, University of Utah, Salt Lake City, USA
  • Zhang, Min-Min Department of Biology, University of Utah, Salt Lake City, USA
  • Yoshikami, Doju Department of Biology, University of Utah, Salt Lake City, USA
  • Bulaj, Grzegorz Department of Medicinal Chemistry, University of Utah, Salt Lake City, USA
  • Olivera, Baldomero M. Department of Biology, University of Utah, Salt Lake City, USA
  • Hughes, Kelly T. Département de Médecine, Université de Fribourg, Switzerland - Department of Biology, University of Utah, Salt Lake City, USA
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    29.05.2012
Published in:
  • mBio. - 2012, vol. 3, no. 3, p. e00115-12t
English The structure, assembly, and function of the bacterial flagellum involves about 60 different proteins, many of which are selectively secreted via a specific type III secretion system (T3SS) (J. Frye et al., J. Bacteriol. 188:2233–2243, 2006). The T3SS is reported to secrete proteins at rates of up to 10,000 amino acid residues per second. In this work, we showed that the flagellar T3SS of Salmonella enterica serovar Typhimurium could be manipulated to export recombinant nonflagellar proteins through the flagellum and into the surrounding medium. We translationally fused various neuroactive peptides and proteins from snails, spiders, snakes, sea anemone, and bacteria to the flagellar secretion substrate FlgM. We found that all tested peptides of various sizes were secreted via the bacterial flagellar T3SS. We subsequently purified the recombinant μ-conotoxin SIIIA (rSIIIA) from Conus striatus by affinity chromatography and confirmed that T3SS-derived rSIIIA inhibited mammalian voltage-gated sodium channel NaV1.2 comparably to chemically synthesized SIIIA.
Faculty
Faculté des sciences et de médecine
Department
Département de Médecine
Language
  • English
Classification
Biology
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License
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Identifiers
  • RERO DOC 29590
Persistent URL
https://folia.unifr.ch/unifr/documents/302477
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