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  Backbone dynamics of green fluorescent protein and the effect of histidine 148 substitution

Seifert, M. H. J., Georgescu, J., Ksiazek, D., Smialowski, P., Rehm, T., Steipe, B., et al. (2003). Backbone dynamics of green fluorescent protein and the effect of histidine 148 substitution. Biochemistry, 42(9), 2500-2512.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-6C71-2 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-6C72-F
Genre: Journal Article
Alternative Title : Biochemistry

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 Creators:
Seifert, M. H. J.1, Author              
Georgescu, J.2, Author              
Ksiazek, D., Author
Smialowski, P.3, 4, Author              
Rehm, T.1, Author              
Steipe, B., Author
Holak, T. A.2, Author              
Affiliations:
1External Organizations, ou_persistent22              
2Holak, Tad / NMR Spectroscopy, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565154              
3Huber, Robert / Structure Research, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565155              
4Conti, Elena / Structural Cell Biology, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565144              

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 Abstract: Green fluorescent protein (GFP) and its mutants have become valuable tools in molecular biology. GFP has been regarded as a very stable and rigid protein with the beta-barrel shielding the chromophore from the solvent. Here, we report the N-15 nuclear magnetic resonance (NMR) studies on the green fluorescent protein (GFPuv) and its mutant His148Gly. N-15 NMR relaxation studies of GFPuv show that most of the beta-barrel of GFP is rigid on the picosecond to nanosecond time scale. For several regions, including the first alpha-helix and beta- sheets 3, 7, 8, and 10, increased hydrogen-deuterium exchange rates suggest a substantial conformational flexibility on the microsecond to millisecond time scales. Mutation of residue 148 located in beta-sheet 7 is known to have a strong impact on the fluorescence properties of GFPs. UV absorption and fluorescence spectra in combination with H-1-N-15 NMR spectra indicate that the His148Gly mutation not only reduces the absorption of the anionic chromophore state but also affects the conformational stability, leading to the appearance of doubled backbone amide resonances for a number of residues. This suggests the presence of two conformations in slow exchange on the NMR time scale in this mutant.

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Language(s): eng - English
 Dates: 2003-03-11
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 41341
ISI: 000181372200002
 Degree: -

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Title: Biochemistry
  Alternative Title : Biochemistry
Source Genre: Journal
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Pages: - Volume / Issue: 42 (9) Sequence Number: - Start / End Page: 2500 - 2512 Identifier: ISSN: 0006-2960