title: Purification and characterisation of recombinant His-tagged RgpB gingipain from Porphymonas gingivalis.
abstract: Gingipain proteases are important virulence factors from the periodontal pathogen Porphyromonas gingivalis and are the target of many in vitro studies. Due to their close biochemical properties, purification of individual gingipains is difficult and requires multiple chromatographic steps. In this study, we demonstrate that insertion of a hexahistidine affinity tag upstream of a C-terminal outer membrane translocation signal in RgpB gingipain leads to the secretion of a soluble, mature form of RgpB bearing the affinity tag that can easily be purified by nickel-chelating affinity chromatography. The final product obtained high yielding high purity is biochemically indistinguishable from the native RgpB enzyme.
authors.forename: Florian
authors.initials: F
authors.lastname: Veillard
authors.forename: Barbara
authors.initials: B
authors.lastname: Potempa
authors.forename: Yonghua
authors.initials: Y
authors.lastname: Guo
authors.forename: Miroslaw
authors.initials: M
authors.lastname: Ksiazek
authors.forename: Maryta N
authors.initials: MN
authors.lastname: Sztukowska
authors.forename: John A
authors.initials: JA
authors.lastname: Houston
authors.forename: Lahari
authors.initials: L
authors.lastname: Koneru
authors.forename: Ky-Anh
authors.initials: KA
authors.lastname: Nguyen
authors.forename: Jan
authors.initials: J
authors.lastname: Potempa
mesh_terms: D018829:Adhesins, Bacterial / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D016866:Bacteroidaceae Infections / Q000382:microbiology; D002846:Chromatography, Affinity; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D000080867:Gingipain Cysteine Endopeptidases; D006801:Humans; D016966:Porphyromonas gingivalis / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D018829:Adhesins, Bacterial; D000080867:Gingipain Cysteine Endopeptidases; D011994:Recombinant Proteins; D003546:Cysteine Endopeptidases