Protein crystallization facilitated by molecularly imprinted polymers
Saridakis, E, Khurshid, S, Govada, L, Phan, Q, Hawkins, D, Crichlow, GV, Lolis, E, Reddy, SM and Chayen, NE (2011) Protein crystallization facilitated by molecularly imprinted polymers Proceedings of the National Academy of Sciences USA, 108 (27). pp. 11081-11086.
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We present a new initiative and its application, namely the design of molecularly imprinted polymers (MIPs) for producing protein crystals which are essential for determining high-resolution 3-D structures of proteins. MIPs, also referred to as ‘smart materials’ are made to contain cavities capable of rebinding protein, thus the fingerprint of the protein created on the polymer allows it to serve as an ideal template for crystal formation. We have shown that six different MIPs induced crystallization of nine proteins, yielding crystals in conditions that do not give crystals otherwise. The incorporation of MIPs in screening experiments gave rise to crystalline hits in 8- 10% of the trials for three target proteins. These hits would have been missed using other known nucleants. MIPs also facilitated the formation of large single crystals at metastable conditions for seven proteins. Moreover, the presence of MIPs has led to faster formation of crystals in all cases where crystals would appear eventually and to major improvement in diffraction in some cases. The MIPs were effective for their cognate proteins and also for other proteins, with size-compatibility being a likely criterion for efficacy. Atomic Force Microscopy (AFM) measurements demonstrated specific affinity between the MIPs cavities and a protein-functionalised AFM tip, corroborating our hypothesis that due to the recognition of proteins by the cavities, MIPs can act as nucleation inducing substrates (nucleants) by harnessing the proteins themselves as templates.
|Divisions :||Faculty of Engineering and Physical Sciences > Chemistry|
|Date :||5 July 2011|
|Identification Number :||10.1073/pnas.1016539108|
|Projects :||EP/G014299/1, EP/G014736/1|
|Uncontrolled Keywords :||crystallography, critical nucleus, proteomics, macromolecules, vapor diffusion, HETEROGENEOUS NUCLEATION, CRYSTAL NUCLEATION, RECOGNITION, PHASE, SUBSTRATE, SURFACES, CHROMATOGRAPHY, DERIVATIVES, TEMPLATE|
|Depositing User :||Symplectic Elements|
|Date Deposited :||12 Oct 2011 10:53|
|Last Modified :||23 Sep 2013 18:46|
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