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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Refined crystal structure of DsRed, a red fluorescent protein from coral, at 2.0-A resolution
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Refined crystal structure of DsRed, a red fluorescent protein from coral, at 2.0-A resolution

机译：DsRed（珊瑚红色荧光蛋白）的精细晶体结构，分辨率为2.0-A

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摘要

The crystal Structure of DsRed, a red fluorescent protein from a corallimorpharian, has been determined at 2.0-A resolution by multiple-wavelength anomalous dispersion and crystallographic refinement. Crystals of the selenomathionine-substituted protein have space group P21 and contain a tetramer with 222 noncrystallo- graphic symmetry in the asymmetric unit. The refined model has satisfactory stereochemistry and a final crystallographi(R factor of 0.162. The protein, which forms an obligatory tetramer in solution and in the crystal, is a squat rectangular prism comprising four protomers whose fold is extremely similar to that of the Aequorea victoria green fluorescent protein despite low (≈23/100) amino add sequence homology. The monomer consists of an 11-stranded p barrel with a coaxial helix. The chromophores, formed from the primary sequence -Gln-Tyr-Gly- (residues 66--68). are arranged in a ≈27 × 34-A rectangular array in two approximately antiparallel pairs. The geometry at the o carbon of Gln-66 (refined without stereochem- ical restraints) is consistent with an sp~2 hybridized center, in accord with the proposal that red fluorescence is because of an additional oxidation step that forms an acylimine extension to the chromophore [Gross, L. A., Baird, G. S., Hoffman. R. C., Baldridge. K. K. & Tsien, R. Y. (2000) Proc. Natl. Acad. Sci. USA 87, 11990-11995]. The carbonyl oxygen of Phe-65 is almost 90° out of the plane of the chromophore, consistent with theoretical calculations suggesting that this is the minimum energy con formation of this moiety despite the conjuga- tion of this group with the rest of the chromophore.
机译：通过多波长异常色散和晶体学细化，以2.0-A的分辨率确定了DsRed的晶体结构，它是一种来自珊瑚形藻的红色荧光蛋白。硒代甲硫氨酸取代蛋白的晶体具有空间群P21，并在不对称单元中包含具有222个非晶体对称性的四聚体。精炼的模型具有令人满意的立体化学和最终的结晶学系数（R因子为0.162。该蛋白质在溶液中和晶体中形成一种必不可少的四聚体）是一个蹲下的直角棱柱，由四个原角组成，其折叠与维多利亚水母非常相似。尽管氨基添加序列同源性较低（≈23/ 100），但仍为绿色荧光蛋白。该单体由11链p桶和同轴螺旋组成。发色团由一级序列-Gln-Tyr-Gly-（残基66- -68）。以≈27×34-A矩形阵列的形式排列，成两对近似反平行的对。Gln-66的o碳原子处的几何形状（无立体化学约束精制）与sp〜2杂交中心一致，根据红色荧光的提议是由于额外的氧化步骤，该步骤形成了向发色团的酰基亚胺延伸[Gross，LA，Baird，GS，Hoffman.RC，Baldridge.KK＆Tsien，RY（2000）Proc.Natl.Acad.Sci.USA，88：3587-2877]。科学院USA 87，11990-11995]。 Phe-65的羰基氧几乎偏离发色团平面90°，这与理论计算相符，这表明尽管该基团与发色团的其余部分共轭，但这是该部分的最小能量构象。

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