Improved protein-crystal identification by using 2,2,2-trichloroethanol as a 
fluorescence enhancer

Pichlo, Christian; Toelzer, Christine; Chojnacki, Konrad; Oecal, Sinan; Uthoff, Matthias; Ruegenberg, Sabine; Hermanns, Thomas; Schacherl, Magdalena; Denzel, Martin S.; Hofmann, Kay; Niefind, Karsten; Baumann, Ulrich

doi:10.1107/S2053230X18005253

Improved protein-crystal identification by using 2,2,2-trichloroethanol as a fluorescence enhancer

Pichlo, C., Toelzer, C., Chojnacki, K., Oecal, S., Uthoff, M., Ruegenberg, S., Hermanns, T., Schacherl, M., Denzel, M. S., Hofmann, K., Niefind, K., & Baumann, U. (2018). Improved protein-crystal identification by using 2,2,2-trichloroethanol as a fluorescence enhancer. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, 74(Pt 5), 307-314. doi:10.1107/S2053230X18005253.

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アイテムのパーマリンク: https://hdl.handle.net/21.11116/0000-0003-53AB-F 版のパーマリンク: https://hdl.handle.net/21.11116/0000-0008-83BD-F

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作成者:
Pichlo, Christian¹, 著者
Toelzer, Christine¹, 著者
Chojnacki, Konrad¹, 著者
Oecal, Sinan¹, 著者
Uthoff, Matthias¹, 著者
Ruegenberg, Sabine¹, 著者
Hermanns, Thomas¹, 著者
Schacherl, Magdalena², 著者
Denzel, Martin S.¹, 著者
Hofmann, Kay¹, 著者
Niefind, Karsten¹, 著者
Baumann, Ulrich¹, 著者

所属:
1External Organizations, ou_persistent22
2Max Planck Research Group Structural Dynamics of Proteins, Center of Advanced European Studies and Research (caesar), Max Planck Society, Ludwig-Erhard-Allee 2, 53175 Bonn, DE, ou_2173687

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キーワード: protein-crystal identification; tryptophan fluorescence; 2,2,2-trichloroethanol; PPEP-1

要旨: The identification of initial lead conditions for successful protein crystallization is crucial for structural studies using X-ray crystallography. In order to reduce the number of false-negative conditions, an emerging number of fluorescence-based methods have been developed which allow more efficient identification of protein crystals and help to distinguish them from salt crystals. Detection of the native tryptophan fluorescence of protein crystals is one of the most widely used methods. However, this method can fail owing to the properties of the crystallized protein or the chemical composition of the crystallization trials. Here, a simple, fast and cost-efficient method employing 2,2,2-trichloroethanol (TCE) has been developed. It can be performed with a standard UV-light microscope and can be applied to cases in which detection of native tryptophan fluorescence fails. In four test cases this method had no effect on the diffraction properties of the crystals and no structural changes were observed. Further evidence is provided that TCE can be added to crystallization trials during their preparation, making this method compatible with high-throughput approaches.

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言語: eng - English

日付: 出版: 2018-05-01

出版の状態: 出版

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査読: 査読あり

識別子（DOI, ISBNなど）: ISI: 000431249400007
DOI: 10.1107/S2053230X18005253

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出版物名: Acta Crystallographica Section F: Structural Biology and Crystallization Communications

省略形 : Acta Crystallogr F Struct Biol Commun

種別: 学術雑誌

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出版社, 出版地: Blackwell Publishing Limited

ページ: - 巻号: 74 (Pt 5) 通巻号: - 開始・終了ページ: 307 - 314 識別子（ISBN, ISSN, DOIなど）: ISSN: 1744-3091
CoNE: https://pure.mpg.de/cone/journals/resource/1000000000017210_1

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