title: Bacterial expression of human vitamin D-binding protein (Gc2) in functional form.
abstract: In this report, we report the first expression of human vitamin D-binding protein (hDBP), a serum protein with several functions and a multidomained structure, in Escherichia coli. The recombinant protein (reDBP) was expressed as a fusion partner of glutathione S-transferase in order to facilitate proper folding of the reDBP; E. coli-expressed DBP was found to be fully functional with respect to vitamin D sterol binding, interaction with actin, and cross-reactivity with anti-DBP antibody. Furthermore, both natural DBP and reDBP were affinity-labeled with 25-hydroxyvitamin D3-3-bromo[1-14C]acetate in a similar fashion. Availability of an expression system for hDBP in functional form provides opportunity to develop mutants and truncated DBPs to study multiple ligand-binding properties of this protein in relationship with its structure.
authors.affiliation: Department of Bioorganic and Protein Chemistry, Boston University School of Medicine, Massachusetts 02118, USA.
authors.forename: N
authors.initials: N
authors.lastname: Swamy
authors.forename: S
authors.initials: S
authors.lastname: Ghosh
authors.forename: R
authors.initials: R
authors.lastname: Ray
mesh_terms: D000199:Actins / Q000378:metabolism; D000345:Affinity Labels; D000595:Amino Acid Sequence; D002117:Calcitriol / Q000378:metabolism; D002846:Chromatography, Affinity; D004926:Escherichia coli; D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D011485:Protein Binding; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000378:metabolism; D013329:Structure-Activity Relationship; D014807:Vitamin D / Q000378:metabolism*; D014809:Vitamin D-Binding Protein / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
chemical_list: D000199:Actins; D000345:Affinity Labels; D011993:Recombinant Fusion Proteins; D014809:Vitamin D-Binding Protein; D014807:Vitamin D; D005982:Glutathione Transferase; D002117:Calcitriol