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Purification of normal cellular prion protein from human platelets and the formation of a high molecular weight prion protein complex following platelet activation

Jones, Michael and Head, Mark W. and Connolly, John and Farquhar, Christine F. and Hornsey, Valerie S. and Pepper, Duncan S. and MacGregor, Ian R. (2005) Purification of normal cellular prion protein from human platelets and the formation of a high molecular weight prion protein complex following platelet activation. Biochemical and Biophysical Research Communications, 335 (1). pp. 48-56. ISSN 1090-2104

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Abstract

A method for the extraction and purification of PrPC, in its native monomeric form, from outdated human platelet concentrates is described. Both calcium ionophore platelet activation and lysis in Triton X-100 were evaluated as methods for the extraction of soluble platelet PrPC in its monomeric form. Following platelet activation, the majority of released PrPC was detected as a disulphide linked high molecular weight complex, which under reducing conditions could be separated into what appear to be stable non-disulphide linked PrP dimers or PrP covalently linked to another as yet unidentified protein. This phenomenon appears to be unique to activation since only monomeric PrPC was detected following lysis of resting platelets. Subsequently, PrPC was purified from the Triton X-100 lysate by sequential cation ion exchange and Cu2+ affinity chromatography. From 10 L of outdated platelet concentrate, we were able to recover 1.29 mg PrPC at a purity of 92%.

Item type: Article
ID code: 37795
Keywords: human prion protein, purification, blood, platelet, calcium ionophore activation, Creutzfeldt-Jakob disease, copper affinity, conformation change, detergent lysis, dimer, cation exchange, Pharmacy and materia medica, Biochemistry, Cell Biology, Molecular Biology, Biophysics
Subjects: Medicine > Pharmacy and materia medica
Department: Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences
Related URLs:
    Depositing user: Pure Administrator
    Date Deposited: 22 Feb 2012 11:52
    Last modified: 04 Sep 2014 19:46
    URI: http://strathprints.strath.ac.uk/id/eprint/37795

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