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Title: Crystallization Optimum Solubility Screening: using crystallization results to identify the optimal buffer for protein crystal formation

Abstract

It is shown how protein crystallization results can be used to identify buffers that improve protein solubility and, in turn, crystallization success. An optimal solubility screen is described that uses the results of crystallization trials to identify buffers that improve protein solubility and, in turn, crystallization success. This screen is useful not only for standard crystallization experiments, but also can easily be implemented into any high-throughput structure-determination pipeline. As a proof of principle, the predicted novel-fold protein AF2059 from Archaeoglobus fulgidus, which was known to precipitate in most buffers and particularly during concentration experiments, was selected. Using the crystallization results of 192 independent crystallization trials, it was possible to identify a buffer containing 100 mM CHES pH 9.25 that significantly improves its solubility. After transferring AF2059 into this ‘optimum-solubility’ buffer, the protein was rescreened for crystal formation against these same 192 conditions. Instead of extensive precipitation, as observed initially, it was found that 24 separate conditions produced crystals and the exchange of AF2059 into CHES buffer significantly improved crystallization success. Fine-screen optimization of these conditions led to the production of a crystal suitable for high-resolution (2.2 Å) structure determination.

Authors:
; ;  [1];  [2]
  1. Department of Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037 (United States)
  2. (United States)
Publication Date:
OSTI Identifier:
22356197
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 61; Journal Issue: Pt 12; Other Information: PMCID: PMC1978149; PMID: 16511228; PUBLISHER-ID: bw5115; OAI: oai:pubmedcentral.nih.gov:1978149; Copyright (c) International Union of Crystallography 2005; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; BUFFERS; CRYSTALLIZATION; CRYSTALS; OPTIMIZATION; PRECIPITATION; PROTEINS; RESOLUTION; SCREENING; SOLUBILITY

Citation Formats

Collins, Bernard, Stevens, Raymond C., Page, Rebecca, E-mail: rebecca-page@brown.edu, and Joint Center for Structural Genomics, 10550 North Torrey Pines Road, La Jolla, CA 92037. Crystallization Optimum Solubility Screening: using crystallization results to identify the optimal buffer for protein crystal formation. United Kingdom: N. p., 2005. Web. doi:10.1107/S1744309105035244.
Collins, Bernard, Stevens, Raymond C., Page, Rebecca, E-mail: rebecca-page@brown.edu, & Joint Center for Structural Genomics, 10550 North Torrey Pines Road, La Jolla, CA 92037. Crystallization Optimum Solubility Screening: using crystallization results to identify the optimal buffer for protein crystal formation. United Kingdom. doi:10.1107/S1744309105035244.
Collins, Bernard, Stevens, Raymond C., Page, Rebecca, E-mail: rebecca-page@brown.edu, and Joint Center for Structural Genomics, 10550 North Torrey Pines Road, La Jolla, CA 92037. Thu . "Crystallization Optimum Solubility Screening: using crystallization results to identify the optimal buffer for protein crystal formation". United Kingdom. doi:10.1107/S1744309105035244.
@article{osti_22356197,
title = {Crystallization Optimum Solubility Screening: using crystallization results to identify the optimal buffer for protein crystal formation},
author = {Collins, Bernard and Stevens, Raymond C. and Page, Rebecca, E-mail: rebecca-page@brown.edu and Joint Center for Structural Genomics, 10550 North Torrey Pines Road, La Jolla, CA 92037},
abstractNote = {It is shown how protein crystallization results can be used to identify buffers that improve protein solubility and, in turn, crystallization success. An optimal solubility screen is described that uses the results of crystallization trials to identify buffers that improve protein solubility and, in turn, crystallization success. This screen is useful not only for standard crystallization experiments, but also can easily be implemented into any high-throughput structure-determination pipeline. As a proof of principle, the predicted novel-fold protein AF2059 from Archaeoglobus fulgidus, which was known to precipitate in most buffers and particularly during concentration experiments, was selected. Using the crystallization results of 192 independent crystallization trials, it was possible to identify a buffer containing 100 mM CHES pH 9.25 that significantly improves its solubility. After transferring AF2059 into this ‘optimum-solubility’ buffer, the protein was rescreened for crystal formation against these same 192 conditions. Instead of extensive precipitation, as observed initially, it was found that 24 separate conditions produced crystals and the exchange of AF2059 into CHES buffer significantly improved crystallization success. Fine-screen optimization of these conditions led to the production of a crystal suitable for high-resolution (2.2 Å) structure determination.},
doi = {10.1107/S1744309105035244},
journal = {Acta Crystallographica. Section F},
number = Pt 12,
volume = 61,
place = {United Kingdom},
year = {Thu Dec 01 00:00:00 EST 2005},
month = {Thu Dec 01 00:00:00 EST 2005}
}
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