XAFS study of the high-affinity copper-binding site of human PrP 91–231 and its low-resolution structure in solution 1 1Edited by I. A. Wilson

Hasnain, S. Samar; Murphy, Loretta M.; Strange, R.; Grossmann, J. Günter; Clarke, AR; Jackson, Graham S.; Collinge, John

doi:10.1006/jmbi.2001.4795

Cited by 125 publications

(91 citation statements)

References 25 publications

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“…Taken together, these findings point toward a sequential model of Cu 2+ uptake in which the higher affinity at His96 site takes up the first copper ion and then facilitates cooperative uptake in the adjacent octarepeat domain. (38). On the basis of the 1:1 Cu 2+ /PrP(91-231) stoichiometry, they proposed that the histidine coordination to the single copper must result from two imidazoles, His96 and His111.…”

Section: Discussionmentioning

confidence: 99%

Copper Coordination in the Full-Length, Recombinant Prion Protein

et al. 2003

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The prion protein (PrP) binds divalent copper at physiologically relevant conditions and is believed to participate in copper regulation or act as a copper-dependent enzyme. Ongoing studies aim at determining the molecular features of the copper binding sites. The emerging consensus is that most copper binds in the octarepeat domain, which is composed of four or more copies of the fundamental sequence PHGGGWGQ. Previous work from our laboratory using PrP-derived peptides, in conjunction with EPR and X-ray crystallography, demonstrated that the HGGGW segment Copper coordination arises from the His imidazole and sequential deprotonated glycine amides. In this present work, recombinant, full-length Syrian hamster PrP is investigated using EPR methodologies. Four copper ions are taken up in the octarepeat domain, which supports previous findings. However, quantification studies reveal a fifth binding site in the flexible region between the octarepeats and the PrP globular C-terminal domain. A series of PrP peptide constructs show that this site involves His96 in the PrP(92-96) segment GGGTH. Further examination by X-band EPR, S-band EPR, and electron spin-echo envelope spectroscopy, demonstrates coordination by the His96 imidazole and the glycine preceding the threonine. The copper affinity for this type of binding site is highly pH dependent, and EPR studies here show that recombinant PrP loses its affinity for copper below pH 6.0. These studies seem to provide a complete profile of the copper binding sites in PrP and support the hypothesis that PrP function is related to its ability to bind copper in a pH-dependent fashion.Prion diseases are fatal neurodegenerative disorders of both humans and animals (1). The causative agent is an isoform of a normal, host-encoded membrane glycoprotein called the prion protein (PrP). 1 The normal cellular isoform (PrP C ) is the precursor to the pathogenic, protease-resistant isoform termed PrP Sc , which is responsible for homologous pathologies within its individual hosts. With rare but notable exceptions, prion diseases respect the species barrier (2,3). Among these exceptions is the transmission of disease from scrapie-infected † This work was supported by NIH Grants GM 65790 (G.L.M.), GM 60609 (G.J.G.), GM 40168 (J.P.), AG02132 and AG10770 (S.B.P.).

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Section: Discussionmentioning

confidence: 99%

Copper Coordination in the Full-Length, Recombinant Prion Protein

et al. 2003

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“…They may arise through a decreased copper concentration in synaptic membranes of PrP Ϫ/Ϫ mice (22). In vitro PrP c has been shown to bind this metal ion at the N-terminal octapeptide repeat regions (6) and within the C-terminal portion of the molecule (21). Alternatively, the absence of PrP c may perturb the regulation of neuronal oxidative stress in which PrP has been implicated through a proposed superoxide dismutase activity (7).…”

Section: Prpmentioning

confidence: 99%

PrP ^c Expression Influences the Establishment of Herpes Simplex Virus Type 1 Latency

Thackray

Bujdoso

2002

J Virol

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Section: Introductionmentioning

confidence: 99%

“…The disease-associated conformer, termed PrP Sc , is a structurally altered form of the normal cellular protein, PrP C . PrP C is a glycosylphosphatidylinositol (GPI)-anchored cell surface glycoprotein, which has been shown to bind copper within an N-terminal octameric repeat region and also at one or more sites slightly more C-terminal of the repeat region [10][11][12][13].Despite much investigation and characterization of the properties of PrP C and PrP Sc , the primary function of PrP C and the principal pathogenic pathways of TSEs remain unresolved. One suggested role for PrP C is the transduction of signals from the external environment into the cell interior, a function circumstantially supported by the localization of PrP C within plasma membrane detergentresistant microdomains (also known as lipid rafts), which are now widely recognized as membrane-signaling plat-…”

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confidence: 99%

PrPC-related signal transduction is influenced by copper, membrane integrity and the alpha cleavage site

et al. 2009

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The copper-binding, membrane-anchored, cellular prion protein (PrP C ) has two constitutive cleavage sites producing distinct N-and C-terminal fragments (N1/C1 and N2/C2). Using RK13 cells expressing either human PrP C , mouse PrP C or mouse PrP C carrying the 3F4 epitope, this study explored the influence of the PrP C primary sequence on endoproteolytic cleavage and one putative PrP C function, MAP kinase signal transduction, in response to exogenous copper with or without a perturbed membrane environment. PrP C primary sequence, especially that around the N1/C1 cleavage site, appeared to influence basal levels of proteolysis at this location and extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation, with increased processing demonstrating an inverse relationship with basal ERK1/2 activation. Human PrP C showed increased N1/C1 cleavage in response to copper alone, accompanied by specific p38 and JNK/SAPK phosphorylation. Combined exposure to copper plus the cholesterol-sequestering antibiotic filipin resulted in a mouse PrP C -specific substantial increase in signal protein phosphorylation, accompanied by an increase in N1/C1 cleavage. Mouse PrP C harboring the human N1/C1 cleavage site assumed more human-like profiles basally and in response to copper and altered membrane environments. Our results demonstrate that the PrP C primary sequence around the N1/C1 cleavage site influences endoproteolytic processing at this location, which appears linked to MAP kinase signal transduction both basally and in response to copper. Further, the primary sequence appears to confer a mutual dependence of N1/C1 cleavage and membrane integrity on the fidelity of PrP C -related signal transduction in response to exogenous stimuli. IntroductionCumulative experimental evidence supports that abnormal isomers of the prion protein constitute the causative agent in transmissible spongiform encephalopathies (TSEs, also known as prion diseases) [1,2], with neuronal expression and membrane anchorage of wild-type PrP being essential for efficient pathogenesis [3][4][5][6][7][8][9]. The disease-associated conformer, termed PrP Sc , is a structurally altered form of the normal cellular protein, PrP C . PrP C is a glycosylphosphatidylinositol (GPI)-anchored cell surface glycoprotein, which has been shown to bind copper within an N-terminal octameric repeat region and also at one or more sites slightly more C-terminal of the repeat region [10][11][12][13].Despite much investigation and characterization of the properties of PrP C and PrP Sc , the primary function of PrP C and the principal pathogenic pathways of TSEs remain unresolved. One suggested role for PrP C is the transduction of signals from the external environment into the cell interior, a function circumstantially supported by the localization of PrP C within plasma membrane detergentresistant microdomains (also known as lipid rafts), which are now widely recognized as membrane-signaling plat-forms [14]. The plasma membrane constitutes a critical site for the activatio...

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XAFS study of the high-affinity copper-binding site of human PrP 91–231 and its low-resolution structure in solution 1 1Edited by I. A. Wilson

Cited by 125 publications

References 25 publications

Copper Coordination in the Full-Length, Recombinant Prion Protein

Copper Coordination in the Full-Length, Recombinant Prion Protein

PrP ^c Expression Influences the Establishment of Herpes Simplex Virus Type 1 Latency

PrPC-related signal transduction is influenced by copper, membrane integrity and the alpha cleavage site

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XAFS study of the high-affinity copper-binding site of human PrP 91–231 and its low-resolution structure in solution 1 1Edited by I. A. Wilson

Cited by 125 publications

References 25 publications

Copper Coordination in the Full-Length, Recombinant Prion Protein

Copper Coordination in the Full-Length, Recombinant Prion Protein

PrP c Expression Influences the Establishment of Herpes Simplex Virus Type 1 Latency

PrPC-related signal transduction is influenced by copper, membrane integrity and the alpha cleavage site

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PrP ^c Expression Influences the Establishment of Herpes Simplex Virus Type 1 Latency