Expression and purification of his-tagged recombinant mouse zeta-crystallin.
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.
Simpanya
Mukoma F
MF
Oakland University, Eye Research Institute, 409 Dodge Hall, Rochester, MI 48309-4480, USA. simpanya@gmail.com
Leverenz
Victor R
VR
Giblin
Frank J
FJ
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*
D011994:Recombinant Proteins; D038226:zeta-Crystallins; D009249:NADP; D011808:Quinone Reductases
Curr Eye Res. 1987 May;6(5):725-32
3595182
Gene. 1989 May 30;78(2):215-24
2777081
J Biol Chem. 1996 Dec 13;271(50):31973-80
8943244
Curr Eye Res. 1982;2(1):39-45
7128182
Exp Eye Res. 2002 Oct;75(4):445-58
12387792
Biochemistry. 1997 May 6;36(18):5353-62
9154917
Cell Biol Int. 1998;22(3):217-25
9974216
J Biol Chem. 2000 Mar 3;275(9):6664-72
10692476
Arch Biochem Biophys. 1996 Apr 1;328(1):173-83
8638928
J Biol Chem. 1995 Nov 3;270(44):26224-31
7592828
Arch Biochem Biophys. 1991 Feb 15;285(1):134-6
1990971
Nucleic Acids Res. 2006;34(14):4060-8
16914438
Nature. 1974 Jul 19;250(463):194-9
4368490
J Biol Chem. 2001 Jul 27;276(30):28134-9
11371565
Cell Mol Life Sci. 2007 Jun;64(11):1419-27
17497241
Invest Ophthalmol Vis Sci. 2007 Apr;48(4):1718-23
17389504
Arch Biochem Biophys. 1991 Jan;284(1):181-5
1989495
Protein Expr Purif. 2001 Mar;21(2):260-7
11237687
Biochim Biophys Acta. 1992 Oct 13;1180(1):44-52
1390943
J Biol Chem. 1992 Jan 5;267(1):96-102
1370456
Biochem Biophys Res Commun. 1993 Mar 31;191(3):902-7
8466529
Invest Ophthalmol Vis Sci. 2005 Dec;46(12):4641-51
16303961
J Mol Biol. 1994 Feb 25;236(3):669-78
8114084
J Biol Chem. 2001 Jun 15;276(24):21375-80
11294877
Mol Biol Evol. 1994 Mar;11(2):305-15
8170370
Photochem Photobiol. 2008 Nov-Dec;84(6):1589-95
18627516