Expression and purification of his-tagged recombinant mouse zeta-crystallin.
Expression and purification of his-tagged recombinant mouse zeta-crystallin.
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DOI：
10.1016/j.pep.2009.08.001
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发表时间：
2010-02
影响因子：
1.6
通讯作者：
Giblin, Frank J.
中科院分区：
文献类型：
--
作者：
Simpanya, Mukoma F.;Leverenz, Victor R.;Giblin, Frank J.
关键词：
β-晶状体蛋白是一种NADPH结合蛋白，由4个相同的35 kD亚基组成。该蛋白质具有醌氧化还原酶活性，大量存在于骆驼科动物、某些雨形啮齿动物和日本树蛙的晶状体中，但在不同物种的某些组织中催化量较低。在这项研究中，重组方法被用来产生大量的组氨酸标记的重组小鼠β-晶状体蛋白，然后将其纯化至同质。重组小鼠β-晶体蛋白的纯化收率是重组豚鼠β-晶体蛋白纯化收率的5倍。纯化的组氨酸标记的重组小鼠β-晶状体蛋白的醌氧化还原酶活性与纯化的天然豚鼠透镜β-晶状体蛋白的醌氧化还原酶活性以及先前报道的重组豚鼠β-晶状体蛋白的醌氧化还原酶活性相当。该方法允许生产大量的重组β-晶状体蛋白，用于对这种感兴趣的蛋白质的生物学作用进行研究，该蛋白质以如此高的浓度存在于某些物种的晶状体中。
ζ-crystallin is an NADPH-binding protein consisting of four identical 35 kD 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 ζ-crystallin, which was then purified to homogeneity. The yield of pure recombinant mouse ζ-crystallin was 5 times that obtained previously for purification of recombinant guinea pig ζ-crystallin. The quinone oxidoreductase activity of purified his-tagged recombinant mouse ζ-crystallin was comparable to that of purified native guinea pig lens ζ-crystallin, and to that previously reported for recombinant guinea pig ζ-crystallin. The method permits production of substantial amounts of recombinant ζ-crystallin for conducting studies on the biological role of this interesting protein, which exists in such high concentration in the lenses of certain species.
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影响因子：
3.9
作者：
Edwards, KJ;Barton, JD;Ollis, DL
通讯作者：
Ollis, DL
DOI：
10.1016/0925-4439(92)90025-i
发表时间：
1992-10-13
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
RODRIGUEZ, IR;GONZALEZ, P;BORRAS, T
通讯作者：
BORRAS, T
影响因子：
4.4
作者：
Simpanya, MF;Ansari, RR;Giblin, FJ
通讯作者：
Giblin, FJ
影响因子：
4.8
作者：
BABIYCHUK, E;KUSHNIR, S;INZE, D
通讯作者：
INZE, D
影响因子：
4.8
作者：
Cobb, BA;Petrash, JM
通讯作者：
Petrash, JM