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Addition of magnesium chloride to enhance mono-dispersity of a coiled-coil recombinant mouse macrophage protein

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Abstract

X-ray crystallography for the determination of three-dimensional structures of protein macromolecules represents an important tool in function assignment of uncharacterized proteins. However, crystallisation is often difficult to achieve. A protein sample fully characterized in terms of dispersity may increase the likelihood of successful crystallisation by improving the predictability of the crystallisation process. To maximize the probability of crystallisation of a novel mouse macrophage protein (rMMP), target molecule was characterized and refined to improve monodispersity. Addition of MgCl2 at low concentrations resolves the rMMP into a monodisperse solution, and finally successful crystallization of rMMP was achieved. The effect of MgCl2 was studied using gel filtration chromatography and dynamic light scattering.

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Acknowledgments

Authors thank all members of the University of Queensland, Australia structural genomics group who inspired us to work on this protein. MP thanks Deakin University for encouraging collaboration with participating institutions. PP thanks DU for providing trainee scholarship to pursue partial work at bioprocessing laboratory.

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Authors and Affiliations

  1. Fermentation and Protein Biotechnology Laboratory, Department of Biotechnology, Punjabi University, Patiala, India

    Parveen Pahuja & Munish Puri

  2. Department of Biophysics, All India Institute of Medical Sciences, New Delhi, India

    Alagiri Srinivasan

  3. Centre for Chemistry and Biotechnology, Deakin University, Geelong Technology Precinct, Geelong, VIC, 3217, Australia

    Munish Puri

Authors
  1. Parveen Pahuja
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  2. Alagiri Srinivasan
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  3. Munish Puri
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Correspondence to Munish Puri.

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Pahuja, P., Srinivasan, A. & Puri, M. Addition of magnesium chloride to enhance mono-dispersity of a coiled-coil recombinant mouse macrophage protein. Mol Cell Biochem 389, 133–139 (2014). https://doi.org/10.1007/s11010-013-1934-x

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  • DOI: https://doi.org/10.1007/s11010-013-1934-x

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