Metal‐Mediated Self‐Assembly of a β‐Sandwich Protein |
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| Authors: | Peter?B Crowley Dr Pedro?M Matias Dr Amir?R Khan Dr Manfred Roessle Dr Dmitri?I Svergun Dr |
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| Institution: | 1. UCD Centre for Synthesis and Chemical Biology, University College Dublin, Belfield, Dublin 4 (Ireland);2. Present Address: School of Chemistry, National University of Ireland, Galway, University Road, Galway (Ireland);3. Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Apartado 127, 2781‐901 Oeiras (Portugal);4. School of Biochemistry and Immunology, Trinity College, Dublin 2 (Ireland);5. European Molecular Biology Laboratory, Hamburg Outstation, Notkestraβe 85, 22603 Hamburg (Germany);6. Institute of Crystallography, Russian Academy of Sciences, Leninsky pr. 59, 117333 Moscow (Russia) |
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| Abstract: | The β‐sandwich cupredoxin Plastocyanin (Pc) was found to self‐assemble in the presence of Zn2+, a known mediator of protein–protein interfaces. Diffraction‐quality crystals of Pc grew from solutions containing zinc acetate as the sole precipitant. Di‐ and trinuclear zinc sites contribute to the crystal contacts in this structure. A different crystal form, also involving numerous zinc bridging ions, was obtained in the presence of poly(ethylene glycol) 8 000. Comparison of the two crystal forms reveals the effect of macromolecular crowding on self‐assembly. Solution‐state structural characterisation of the Zn2+‐mediated Pc oligomers was performed by using a combination of chemical shift perturbation mapping and small‐angle X‐ray scattering. The data indicate the formation of dimers in solution. The implications for metal‐mediated assembly and crystallisation are discussed. |
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| Keywords: | crystal growth macromolecular crowding methionine protein– protein interactions X‐ray scattering |
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