Efficient production of human beta-defensin 2 (HBD2) in Escherichia coli.
Human beta-defensin 2 (HBD2) has been shown to interact with pathogenic bacteria and components of the mammalian innate and adaptive immune response. We describe a quick and reliable method for the production of HBD2 in Escherichia coli. HBD2 was expressed as an insoluble fusion, chemically cleaved and oxidised to give a single, folded HBD2 beta-isoform. The purified peptide was analysed by high resolution mass spectrometry, displayed a well-dispersed (1)H NMR spectrum, was a chemoattractant to HEK293 cells expressing CCR6 and acted as an antimicrobial agent against E. coli, P. aeruginosa, C. albicans and S. aureus.
Vargues
Thomas
T
School of Chemistry, EastChem, University of Edinburgh, Edinburgh, Scotland, Scotland, United Kingdom.
Morrison
Gareth J
GJ
Seo
Emily S
ES
Clarke
David J
DJ
Fielder
Helen L
HL
Bennani
Julien
J
Pathania
Uday
U
Kilanowski
Fiona
F
Dorin
Julia R
JR
Govan
John R W
JR
Mackay
C Logan
CL
Uhrín
Dusan
D
Campopiano
Dominic J
DJ
D000595:Amino Acid Sequence; D000890:Anti-Infective Agents / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects; D001483:Base Sequence; D002460:Cell Line; D002633:Chemotaxis / Q000187:drug effects; D004926:Escherichia coli / Q000235:genetics*; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D017510:Protein Folding; D054399:Receptors, CCR6 / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D023083:beta-Defensins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology
D000890:Anti-Infective Agents; C106311:CCR6 protein, human; C106641:DEFB4A protein, human; D054399:Receptors, CCR6; D011993:Recombinant Fusion Proteins; D023083:beta-Defensins