2H5R
Crystal structure of mStrawberry at pH 10.5
2H5R の概要
エントリーDOI | 10.2210/pdb2h5r/pdb |
関連するPDBエントリー | 2H5O 2H5P 2H5Q |
分子名称 | mStrawberry (2 entities in total) |
機能のキーワード | beta barrel, luminescent protein |
由来する生物種 | Discosoma sp. |
タンパク質・核酸の鎖数 | 1 |
化学式量合計 | 26675.34 |
構造登録者 | |
主引用文献 | Shu, X.,Shaner, N.C.,Yarbrough, C.A.,Tsien, R.Y.,Remington, S.J. Novel Chromophores and Buried Charges Control Color in mFruits Biochemistry, 45:9639-9647, 2006 Cited by PubMed Abstract: mFruits are second-generation monomeric red fluorescent proteins (mRFPs) that have improved brightness and photostability compared to the first-generation mRFP1. The emission and excitation maxima are distributed over the remarkably large ranges of about 550-650 and 540-590 nm, respectively; however, the variations in the spectra can be traced to a few key amino acids. Spectroscopic and atomic resolution crystallographic analyses of three representatives, mOrange, mStrawberry, and mCherry, reveal that different mechanisms operate to establish the excitation and emission maxima. Evidently, they all undergo the second oxidation step to produce an acylimine linkage in the polypeptide backbone. In comparison to the progenitor DsRed, direct covalent modification to this linkage (mOrange) and indirect modification of the chromophore environment (mStrawberry and mCherry) produce strong blue- and red-shifted variants. The blue shift of mOrange is induced by an unprecedented covalent modification of the protein backbone. The electron-density map indicates the formation of a third heterocycle, 2-hydroxy-dihydrooxazole, upon the reaction of Thr 66 Ogamma with the polypeptide backbone, which in turn reduces the conjugation of the carbonyl at position 65 with the rest of the chromophore. In mStrawberry and mCherry, the movement of charged Lys 70 and protonation of Glu 215 are proposed to modify the chromophore electron-density distribution, inducing the red shift. pH-dependent spectral shifts of mCherry and mStrawberry appear to result from the titration of Glu 215, although, for mStrawberry, partial cyclization of Thr 66 may contribute at high pH. PubMed: 16893165DOI: 10.1021/bi060773l 主引用文献が同じPDBエントリー |
実験手法 | X-RAY DIFFRACTION (1.6 Å) |
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