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Utility of anion and cation combinations for phasing of protein structures

  • Published:
Journal of Structural and Functional Genomics

Abstract

We report the use of anionic (I−), cationic (Ba2+, Cd2+) and ionic mixtures (I− plus Ba2+) for derivatizing liver fatty acid binding protein (LFABP) crystals. Use of cationic and anionic salts in phasing experiments revealed distinct non-overlapping sites for these ions, suggesting exclusive binding regions on LFABP. Interestingly, cations of identical charge and valency (like Ba2+ and Cd2+) bound to distinct pockets on the protein surface. Furthermore, a mixture of salts containing both I− and Ba2+ was very useful in phasing experiments as these oppositely charged ions bound to different regions of LFABP. Our data therefore suggest that cationic and anionic salt mixtures like BaCl2 with NH4I or salts like CdI, BaI where each ion has a significant anomalous signal for a given X-ray wavelength may be valuable reagents for phasing during structure determination.

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Abbreviations

LFABP:

Liver fatty acid binding protein

SAD:

Single wavelength anomalous dispersion

MIR:

Mutiple isomorphous replacement

S-SAD:

Sulphur-SAD

I-SAD:

Iodide-SAD

Ba-SAD:

Barium-SAD

Cd-SAD:

Cadmium-SAD

RMSD:

Root mean square deviation

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Acknowledgments

The X-ray facility at ICGEB was funded by the Wellcome Trust. Ashwani Sharma is supported by University Grants Commission, Government of India. MY is supported by FP7 grants from the EU. Funding for research on “Exploration of salts combinations for fast phasing of protein crystals” is provided by a Department of Biotechnology, Government of India grant to the laboratory.

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Authors and Affiliations

  1. Structural and Computational Biology Group, International Centre for Genetic Engineering and Biotechnology (ICGEB), Aruna Asaf Ali Road, New Delhi, 110067, India

    Ashwani Sharma, Manickam Yogavel & Amit Sharma

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  1. Ashwani Sharma
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  2. Manickam Yogavel
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  3. Amit Sharma
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Corresponding author

Correspondence to Amit Sharma.

Additional information

Ashwani Sharma and Manickam Yogavel contributed equally.

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Sharma, A., Yogavel, M. & Sharma, A. Utility of anion and cation combinations for phasing of protein structures. J Struct Funct Genomics 13, 135–143 (2012). https://doi.org/10.1007/s10969-012-9137-3

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  • DOI: https://doi.org/10.1007/s10969-012-9137-3

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