title: Bacterial expression and refolding of different fragments of human CD14.
abstract: We have investigated bacterial expression of several fragments of CD14, a human cellular receptor for lipopolysaccharides (LPS). Despite binding of CD14 to the LPS, a vital constituent of bacterial outer membrane, we have succeeded in producing full length recombinant hCD14 in E. coli. High level of production of CD14 resulted in deposits of aggregated CD14 in bacteria in form of inclusion bodies, which made production of this protein possible. We have also produced N-terminal fragments consisting of 134 and 152 residues, which comprise N-terminal domain with 2 and 3 leucine rich repeats, respectively and a fragment that contains only leucine rich repeats. Production of the N-terminal domain consisting of 69 residues could not be detected, probably due to the degradation of the produced protein within the bacteria. Full length CD14 and a fragment of 152 residues from inclusion bodies were refolded, while the 134 residues fragment and the one with 10 leucine-rich repeats could not be refolded. Those results confirm that the minimal folding unit of CD14 must include N-terminal domain and at least 3 leucine rich repeats.
authors.affiliation: Laboratory for molecular modeling and NMR spectroscopy, National Institute of Chemistry, Ljubljana, Slovenia.
authors.forename: A
authors.initials: A
authors.lastname: Majerle
authors.forename: J
authors.initials: J
authors.lastname: Kidric
authors.forename: R
authors.initials: R
authors.lastname: Jerala
mesh_terms: D004926:Escherichia coli / Q000378:metabolism*; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017510:Protein Folding; D011994:Recombinant Proteins / Q000378:metabolism; D020449:Repetitive Sequences, Amino Acid
chemical_list: D018950:Lipopolysaccharide Receptors; D010446:Peptide Fragments; D011994:Recombinant Proteins