Universität zu Köln

Pichlo, Christian ORCID: 0000-0002-3156-5667, Toelzer, Christine ORCID: 0000-0001-9984-5940, Chojnacki, Konrad ORCID: 0000-0002-9285-0904, Oecal, Sinan, Uthoff, Matthias ORCID: 0000-0002-6477-5505, Ruegenberg, Sabine, Hermanns, Thomas ORCID: 0000-0001-5350-7479, Schacherl, Magdalena ORCID: 0000-0002-5478-2509, Denzel, Martin S., Hofmann, Kay, Niefind, Karsten ORCID: 0000-0002-0183-6315 and Baumann, Ulrich (2018). Improved protein-crystal identification by using 2,2,2-trichloroethanol as a fluorescence enhancer. Acta Crystallogr. F-Struct. Biol. Commun., 74. S. 307 - 315. CHESTER: INT UNION CRYSTALLOGRAPHY. ISSN 2053-230X

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Abstract

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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Pichlo, Christian UNSPECIFIED orcid.org/0000-0002-3156-5667 UNSPECIFIED Toelzer, Christine UNSPECIFIED orcid.org/0000-0001-9984-5940 UNSPECIFIED Chojnacki, Konrad UNSPECIFIED orcid.org/0000-0002-9285-0904 UNSPECIFIED Oecal, Sinan UNSPECIFIED UNSPECIFIED UNSPECIFIED Uthoff, Matthias UNSPECIFIED orcid.org/0000-0002-6477-5505 UNSPECIFIED Ruegenberg, Sabine UNSPECIFIED UNSPECIFIED UNSPECIFIED Hermanns, Thomas UNSPECIFIED orcid.org/0000-0001-5350-7479 UNSPECIFIED Schacherl, Magdalena UNSPECIFIED orcid.org/0000-0002-5478-2509 UNSPECIFIED Denzel, Martin S. UNSPECIFIED UNSPECIFIED UNSPECIFIED Hofmann, Kay UNSPECIFIED UNSPECIFIED UNSPECIFIED Niefind, Karsten UNSPECIFIED orcid.org/0000-0002-0183-6315 UNSPECIFIED Baumann, Ulrich UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-188384
DOI:	10.1107/S2053230X18005253
Journal or Publication Title:	Acta Crystallogr. F-Struct. Biol. Commun.
Volume:	74
Page Range:	S. 307 - 315
Date:	2018
Publisher:	INT UNION CRYSTALLOGRAPHY
Place of Publication:	CHESTER
ISSN:	2053-230X
Language:	English
Faculty:	Faculty of Mathematics and Natural Sciences
Divisions:	Faculty of Mathematics and Natural Sciences > Department of Biology > Institute for Genetics
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language STRUCTURE REFINEMENT; CRYSTALLIZATION; TRYPTOPHAN; BOND; TOOL Multiple languages Biochemical Research Methods; Biochemistry & Molecular Biology; Biophysics; Crystallography Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/18838