title: Expression and purification of his-tagged recombinant mouse zeta-crystallin.
abstract: Zeta-crystallin is an NADPH-binding protein consisting of four identical 35kD subunits. The protein possesses quinone oxidoreductase activity, and is present in large amounts in the lenses of camelids, certain hystricomorphic rodents, and the Japanese tree frog, and in lower catalytic amounts in certain tissues of various species. In this study, recombinant methods were used to produce substantial quantities of his-tagged recombinant mouse zeta-crystallin, which was then purified to homogeneity. The yield of pure recombinant mouse zeta-crystallin was five times that obtained previously for purification of recombinant guinea pig zeta-crystallin. The quinone oxidoreductase activity of purified his-tagged recombinant mouse zeta-crystallin was comparable to that of purified native guinea pig lens zeta-crystallin, and to that previously reported for recombinant guinea pig zeta-crystallin. The method permits production of substantial amounts of recombinant zeta-crystallin for conducting studies on the biological role of this interesting protein, which exists in such high concentration in the lenses of certain species.
authors.affiliation: Oakland University, Eye Research Institute, 409 Dodge Hall, Rochester, MI 48309-4480, USA. simpanya@gmail.com
authors.forename: Mukoma F
authors.initials: MF
authors.lastname: Simpanya
authors.forename: Victor R
authors.initials: VR
authors.lastname: Leverenz
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Giblin
mesh_terms: D000818:Animals; D006168:Guinea Pigs; D007908:Lens, Crystalline / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D009249:NADP / Q000378:metabolism; D011808:Quinone Reductases / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D038226:zeta-Crystallins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D038226:zeta-Crystallins; D009249:NADP; D011808:Quinone Reductases
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