title: Surface histidine mutations for the metal affinity purification of a β-carbonic anhydrase.
abstract: Metal affinity chromatography using polyhistidine tags is a standard laboratory technique for the general purification of proteins from cellular systems, but there have been no attempts to explore whether the surface character of a protein may be engineered to similar affinity. We present the Arg160His mutation of Haemophilus influenzae carbonic anhydrase (HICA), which mimics the endogenous metal affinity of Escherichia coli carbonic anhydrase (ECCA). The purity and activity of the mutant are reported, and the purification is discussed. This is the first step toward developing a general method to engineer surface metal affinity for use in purification and metal labeling techniques.
authors.affiliation: Department of Chemistry and Biochemistry, Gonzaga University, Spokane, WA 99258, USA. Electronic address: hoffmann@gonzaga.edu.
authors.forename: Katherine M
authors.initials: KM
authors.lastname: Hoffmann
authors.affiliation: Department of Chemistry and Biochemistry, Gonzaga University, Spokane, WA 99258, USA.
authors.forename: Kaitlin M
authors.initials: KM
authors.lastname: Wood
authors.affiliation: Department of Chemistry and Biochemistry, Gonzaga University, Spokane, WA 99258, USA.
authors.forename: Alysha D
authors.initials: AD
authors.lastname: Labrum
authors.affiliation: Department of Chemistry and Biochemistry, Gonzaga University, Spokane, WA 99258, USA.
authors.forename: Dave K
authors.initials: DK
authors.lastname: Lee
authors.affiliation: Department of Chemistry and Biochemistry, Gonzaga University, Spokane, WA 99258, USA.
authors.forename: Ingmar M
authors.initials: IM
authors.lastname: Bolinger
authors.affiliation: Department of Chemistry and Biochemistry, Gonzaga University, Spokane, WA 99258, USA.
authors.forename: Mary E
authors.initials: ME
authors.lastname: Konis
authors.affiliation: Department of Chemistry and Biochemistry, Gonzaga University, Spokane, WA 99258, USA.
authors.forename: Adam G
authors.initials: AG
authors.lastname: Blount
authors.affiliation: Department of Chemistry and Biochemistry, Gonzaga University, Spokane, WA 99258, USA.
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Prussia
authors.affiliation: Department of Chemistry and Biochemistry, Gonzaga University, Spokane, WA 99258, USA.
authors.forename: Monica M
authors.initials: MM
authors.lastname: Schroll
authors.affiliation: Department of Chemistry and Biochemistry, Gonzaga University, Spokane, WA 99258, USA.
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Watson
mesh_terms: D019943:Amino Acid Substitution; D001120:Arginine / Q000378:metabolism; D002256:Carbonic Anhydrases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000201:enzymology; D006193:Haemophilus influenzae / Q000201:enzymology; D006639:Histidine / Q000378:metabolism*; D007700:Kinetics; D013499:Surface Properties
chemical_list: D006639:Histidine; D001120:Arginine; D002256:Carbonic Anhydrases
keywords: Endogenous nickel affinity; Haemophilus influenzae; Immobilized metal affinity purification; Surface histidines; β-Carbonic anhydrase