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Split green fluorescent protein as a modular binding partner for protein crystallization
A modular strategy for protein crystallization using split green fluorescent protein (GFP) as a crystallization partner is demonstrated. Insertion of a hairpin containing GFP β-strands 10 and 11 into a surface loop of a target protein provides two chain crossings between the target and the reconsti...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3852656/ https://www.ncbi.nlm.nih.gov/pubmed/24311592 http://dx.doi.org/10.1107/S0907444913024608 |
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author | Nguyen, Hau B. Hung, Li-Wei Yeates, Todd O. Terwilliger, Thomas C. Waldo, Geoffrey S. |
author_facet | Nguyen, Hau B. Hung, Li-Wei Yeates, Todd O. Terwilliger, Thomas C. Waldo, Geoffrey S. |
author_sort | Nguyen, Hau B. |
collection | PubMed |
description | A modular strategy for protein crystallization using split green fluorescent protein (GFP) as a crystallization partner is demonstrated. Insertion of a hairpin containing GFP β-strands 10 and 11 into a surface loop of a target protein provides two chain crossings between the target and the reconstituted GFP compared with the single connection afforded by terminal GFP fusions. This strategy was tested by inserting this hairpin into a loop of another fluorescent protein, sfCherry. The crystal structure of the sfCherry-GFP(10–11) hairpin in complex with GFP(1–9) was determined at a resolution of 2.6 Å. Analysis of the complex shows that the reconstituted GFP is attached to the target protein (sfCherry) in a structurally ordered way. This work opens the way to rapidly creating crystallization variants by reconstituting a target protein bearing the GFP(10–11) hairpin with a variety of GFP(1–9) mutants engineered for favorable crystallization. |
format | Online Article Text |
id | pubmed-3852656 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-38526562013-12-12 Split green fluorescent protein as a modular binding partner for protein crystallization Nguyen, Hau B. Hung, Li-Wei Yeates, Todd O. Terwilliger, Thomas C. Waldo, Geoffrey S. Acta Crystallogr D Biol Crystallogr Research Papers A modular strategy for protein crystallization using split green fluorescent protein (GFP) as a crystallization partner is demonstrated. Insertion of a hairpin containing GFP β-strands 10 and 11 into a surface loop of a target protein provides two chain crossings between the target and the reconstituted GFP compared with the single connection afforded by terminal GFP fusions. This strategy was tested by inserting this hairpin into a loop of another fluorescent protein, sfCherry. The crystal structure of the sfCherry-GFP(10–11) hairpin in complex with GFP(1–9) was determined at a resolution of 2.6 Å. Analysis of the complex shows that the reconstituted GFP is attached to the target protein (sfCherry) in a structurally ordered way. This work opens the way to rapidly creating crystallization variants by reconstituting a target protein bearing the GFP(10–11) hairpin with a variety of GFP(1–9) mutants engineered for favorable crystallization. International Union of Crystallography 2013-11-19 /pmc/articles/PMC3852656/ /pubmed/24311592 http://dx.doi.org/10.1107/S0907444913024608 Text en © Nguyen et al. 2013 http://creativecommons.org/licenses/by/2.0/uk/ This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited. |
spellingShingle | Research Papers Nguyen, Hau B. Hung, Li-Wei Yeates, Todd O. Terwilliger, Thomas C. Waldo, Geoffrey S. Split green fluorescent protein as a modular binding partner for protein crystallization |
title | Split green fluorescent protein as a modular binding partner for protein crystallization |
title_full | Split green fluorescent protein as a modular binding partner for protein crystallization |
title_fullStr | Split green fluorescent protein as a modular binding partner for protein crystallization |
title_full_unstemmed | Split green fluorescent protein as a modular binding partner for protein crystallization |
title_short | Split green fluorescent protein as a modular binding partner for protein crystallization |
title_sort | split green fluorescent protein as a modular binding partner for protein crystallization |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3852656/ https://www.ncbi.nlm.nih.gov/pubmed/24311592 http://dx.doi.org/10.1107/S0907444913024608 |
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