Purification and characterization of zeta-crystallin/quinone reductase from guinea pig liver.
zeta-Crystallin, a major lens protein of certain mammalian species, has recently been characterized as a novel and active NADPH:quinone oxidoreductase. Here we report the purification of this protein from guinea pig liver by utilizing sequentially: ammonium sulphate precipitation, Blue Sepharose affinity, cation exchange and hydrophobic chromatography steps. This four-step isolation procedure yielded 118-fold purification and a specific activity of 6 U/mg protein when assayed in the presence of 9,10-phenanthrenequinone. Kinetic, immunological and physical properties of this protein have been found to be identical with those of guinea pig lens zeta-crystallin. Western blot analysis using antibodies raised against zeta-crystallin peptides demonstrated the presence of substantial amounts of this protein in human liver homogenates.
Rao
P V
PV
Laboratory of Mechanisms of Ocular Diseases National Eye Institute, National Institutes of Health, Bethesda, MD 20892.
Zigler
J S
JS
D000818:Animals; D001483:Base Sequence; D005591:Chemical Fractionation; D002853:Chromatography, Liquid / Q000379:methods; D003459:Crystallins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006168:Guinea Pigs; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D011808:Quinone Reductases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D003459:Crystallins; D011808:Quinone Reductases