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

Michael Jones; Mark W. Head; John Connolly; Christine F. Farquhar; Valerie S. Hornsey; Duncan S. Pepper; Ian R. MacGregor

doi:10.1016/j.bbrc.2005.07.045

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

Michael Jones, Mark W. Head, John Connolly, Christine F. Farquhar, Valerie S. Hornsey, Duncan S. Pepper, Ian R. MacGregor

Strathclyde Institute Of Pharmacy And Biomedical Sciences

Research output: Contribution to journal › Article › peer-review

9 Citations (Scopus)

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A method for the extraction and purification of PrPC, in its native monomeric form, from outdated human platelet concentrates is described.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 From 10 L of outdated platelet concentrate, we were able to recover 1.29 mg PrPC at a purity of 92%.

Original language	English
Pages (from-to)	48-56
Number of pages	9
Journal	Biochemical and Biophysical Research Communications
Volume	335
Issue number	1
DOIs	https://doi.org/10.1016/j.bbrc.2005.07.045
Publication status	Published - 16 Sep 2005

Keywords

human prion protein
purification
blood
platelet
calcium ionophore activation
Creutzfeldt-Jakob disease
copper affinity
conformation change
detergent lysis
dimer
cation exchange

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.bbrc.2005.07.045

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Jones, M., Head, M. W., Connolly, J., Farquhar, C. F., Hornsey, V. S., Pepper, D. S., & MacGregor, I. 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), 48-56. https://doi.org/10.1016/j.bbrc.2005.07.045

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

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%.",

keywords = "human prion protein, purification, blood, platelet, calcium ionophore activation, Creutzfeldt-Jakob disease, copper affinity, conformation change, detergent lysis, dimer, cation exchange",

author = "Michael Jones and Head, {Mark W.} and John Connolly and Farquhar, {Christine F.} and Hornsey, {Valerie S.} and Pepper, {Duncan S.} and MacGregor, {Ian R.}",

year = "2005",

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doi = "10.1016/j.bbrc.2005.07.045",

language = "English",

volume = "335",

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journal = "Biochemical and Biophysical Research Communications",

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Jones, M, Head, MW, Connolly, J, Farquhar, CF, Hornsey, VS, Pepper, DS & MacGregor, IR 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, vol. 335, no. 1, pp. 48-56. https://doi.org/10.1016/j.bbrc.2005.07.045

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; Head, Mark W.; Connolly, John et al.

In: Biochemical and Biophysical Research Communications, Vol. 335, No. 1, 16.09.2005, p. 48-56.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Purification of normal cellular prion protein from human platelets and the formation of a high molecular weight prion protein complex following platelet activation

AU - Jones, Michael

AU - Head, Mark W.

AU - Connolly, John

AU - Farquhar, Christine F.

AU - Hornsey, Valerie S.

AU - Pepper, Duncan S.

AU - MacGregor, Ian R.

PY - 2005/9/16

Y1 - 2005/9/16

N2 - 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%.

AB - 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%.

KW - human prion protein

KW - purification

KW - blood

KW - platelet

KW - calcium ionophore activation

KW - Creutzfeldt-Jakob disease

KW - copper affinity

KW - conformation change

KW - detergent lysis

KW - dimer

KW - cation exchange

U2 - 10.1016/j.bbrc.2005.07.045

DO - 10.1016/j.bbrc.2005.07.045

M3 - Article

VL - 335

SP - 48

EP - 56

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 1090-2104

IS - 1

ER -

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

Abstract

Keywords

UN SDGs

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