Synthesis and Properties of the Chromophore of the asFP595 Chromoprotein from Anemonia sulcata

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• 作者：Ilia V. Yampolsky ; S. James Remington ; Vladimir I. Martynov ; Victor K. Potapov ; Sergey Lukyanov ; and Konstantin A. Lukyanov
• 刊名：Biochemistry
• 出版年：2005
• 出版时间：April 19, 2005
• 年：2005
• 卷：44
• 期：15
• 页码：5788 - 5793
• 全文大小：189K
• 年卷期：v.44,no.15(April 19, 2005)
• ISSN：1520-4995

文摘

A model compound for the chromophore within the purple nonfluorescent GFP-likechromoprotein asFP595 was synthesized. The postulated structure of the chromophore, 2-acetyl-4-(p-hydroxybenzylidene)-1-methyl-5-imidazolone, was taken from the high-resolution crystal structure analysisof intact asFP595 [Quillin, M. L., Anstrom, D., Shu, X., O'Leary, S., Kallio, K., Lukyanov, K. A., andRemington, S. J. (2005) Kindling Fluorescent Protein from Anemonia sulcata: Dark-State Structure at1.38 Å Resolution, Biochemistry 44, 5774-5787]. Erlenmeyer lactonization and oxidation of the methylenegroup attached to the heteroaromatic moiety with selenium dioxide were used at the key stages of thesynthesis. The spectral properties of the model chromophore in solution and their dependence on the pHand polarity of the solvent were investigated. In water, the chromophore was found to exist in two forms,neutral and anionic, with a pK_a of 7.1. In a dimethylformamide solution, the spectral properties of theanionic form closely match those of the native protein, demonstrating that under these conditions, thecompound is an excellent model for the chromophore within native asFP595.