Cell Surface Accumulation of a Truncated Transmembrane Prion Protein in Gerstmann-Straussler-Scheinker Disease P102L

Mishra, Ravi Shankar; Gu, Yixuan; Bose, Sharmila; Verghese, Susamma; Kalepu, Sudheera; Singh, Neena

doi:10.1074/jbc.m200213200

Cited by 38 publications

(33 citation statements)

References 29 publications

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“…It was surprising to note that a significant amount of PrP C in PT cells was localized to recycling endosomes or late endosomes and lysosomes, a distribution markedly different from the mainly plasma membrane expression on neuronal cells (33,42,43). This can be explained by the large apical endocytosis activity of proximal tubule cells responsible in vivo for the uptake of a large daily load of small filtered proteins (6,8).…”

Section: Discussionmentioning

confidence: 93%

Prion Protein Promotes Kidney Iron Uptake via Its Ferrireductase Activity

Haldar¹,

Tripathi²,

Qian³

et al. 2015

Journal of Biological Chemistry

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Section: Discussionmentioning

confidence: 93%

Prion Protein Promotes Kidney Iron Uptake via Its Ferrireductase Activity

Haldar¹,

Tripathi²,

Qian³

et al. 2015

Journal of Biological Chemistry

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“…The NMR assignments were based on standard triple resonance experiments (36), using a strategy described under "Experimental Procedures." The backbone assignments are complete, with the following exceptions: in all the proteins the H N , 15 N, 13 Screening for a Membrane Mimetic-At the beginning of our study, we tested several detergents and phospholipid bicelles as possible membrane mimetics. The best results were obtained with DPC, whose phosphocholine head groups are expected to simulate a membrane interface, and which is the most widely used detergent for NMR studies of both single ␣-helical membrane proteins and integral membrane ␤-barrel proteins (38,39).…”

Section: Resultsmentioning

confidence: 99%

“…mPrP- were also produced as uniformly 13 C, 15 N-labeled proteins (29,30 15 N, 13 C ␣ , 13 C ␤ , and 13 CO resonance assignments for mPrP-(90 -231) at pH 7.0 were generated by transfer of the published assignments of mPrP-(121-231) at pH 4.5, and of human PrP-(90 -230) at pH 7.0 (29 -31). The assignments were then confirmed using standard triple resonance NMR experiments with the uniformly 13 All NMR experiments were performed at 20°C on a Bruker DRX500 spectrometer equipped with a triply tunable cryogenic probehead. The NMR data were processed with the programs XWINNMR and T0PSPIN 2.0, and the program CARA (www.nmr.ch) was used for the spectral analysis (31).…”

mentioning

confidence: 99%

“…The following mutagenesis primers were used: 5Ј-AGG GGC TGC GGT AGC TGG GGC AG-3Ј and 5Ј-CTG CCC CAG CTA CCG CAG CCC CT-3Ј for mPrP [A117V]-(90 -231), 5Ј-CCA ACC TCA AGC ATG TGG TAG GGG TCG CGG CAG TGG GGG CAG TAG TGG GGG GCC TTG GT-3Ј and 5Ј-ACC AAG GCC CCC CAC TAC TGC CCC CAC TGC CGC GAC CCC TAC CAC ATG CTT GAG GTT GG-3Ј for mPrP[A113V,A115V,A118V]-(90 -231), 5Ј-CCA AAA ACC AAC CTC ATC ATC GTG GCA GGG GCT GCG GCA-3Ј and 5Ј-TGC CGC AGC CCC TGC CAC GAT GAT GAG GTT GGT TTT TGG-3Ј for All the aforementioned constructs were expressed and purified as uniformly 15 N-labeled proteins (29,30). mPrP- were also produced as uniformly 13 C, 15 N-labeled proteins (29,30 15 N, 13 C ␣ , 13 C ␤ , and 13 CO resonance assignments for mPrP-(90 -231) at pH 7.0 were generated by transfer of the published assignments of mPrP-(121-231) at pH 4.5, and of human PrP-(90 -230) at pH 7.0 (29 -31). The assignments were then confirmed using standard triple resonance NMR experiments with the uniformly 13 All NMR experiments were performed at 20°C on a Bruker DRX500 spectrometer equipped with a triply tunable cryogenic probehead.…”

mentioning

confidence: 99%

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Prion Protein-Detergent Micelle Interactions Studied by NMR in Solution

Hornemann

Schroetter

Damberger

et al. 2009

Journal of Biological Chemistry

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Transmissible spongiform encephalopathies (TSEs), 2 such as Creutzfeldt-Jakob disease and the Gerstmann-Sträussler-Scheinker syndrome in humans, are accompanied by the appearance in the brain of an aggregated "scrapie" isoform of the host-encoded prion protein, PrP Sc (1-3). The cellular form, PrP C , consists of an unstructured N-terminal "tail" of residues 23-125 and a globular domain of residues 126 -231, and is attached by a C-terminal glycosylphosphatidylinositol (GPI) anchor to the outer plasma membrane. This structure ensures a role of membrane interactions in the physiological function of PrP C and probably also in the disease-related events leading to TSEs. For example, transgenic mice expressing a prion protein variant lacking the GPI membrane anchor did not develop the typical clinical signs of TSE after inoculation with infectious brain homogenate, although significant amounts of PrP Sc accumulated in the brain (4). This finding led to the conclusion that membrane-association of PrP C is necessary for the development of a TSE. Independent evidence for the importance of membrane interactions for the onset of prion diseases was derived from cell-free conversion assays and cell culture experiments (5, 6).Data have also been presented that indicate that in addition to the normal form with the C terminus linked to a GPI anchor and the C-terminal domain located on the cell surface, PrP C can adopt two different transmembrane topologies, Ctm PrP and Ntm PrP, which have the C-terminal polypeptide segment located in the lumen of the endoplasmic reticulum ( Ctm PrP) or in the cytoplasm ( Ntm PrP) (7-9). The population of the Ctm PrP variant is Ͻ10% of the total wild-type prion protein present during cellular biosynthesis but is increased to 20 -30% for the pathogenic mutations P102L, P105L, and A117V of human PrP and the designed variant mouse PrPs obtained with the amino acid exchanges A113V/A115V/A118V and K110I/H111I (10 -13). The population of Ctm PrP was further increased when an additional mutation, L9R, was present in the N-terminal signal sequence (14), so that ϳ50% of the PrP was synthesized as the Ctm PrP variant in granule neurons obtained from transgenic mice expressing a prion protein construct carrying the four amino acid replacements L9R, A113V, A115V, and A118V (15). Quite generally, an increase in the population of Ctm PrP was also shown to be associated with severe neurodegeneration in transgenic mice, and it has been suggested that Ctm PrP may be the proximate cause of neuronal death in certain prion disorders (10,11,15).In vitro studies on interactions of full-length and N-terminally truncated forms of recombinant PrP showed that acidic membranes caused the N-terminal part of the protein to become more structured, whereas the C-terminal domain was destabilized (16 -19). Furthermore, zwitterionic gel-phase dipalmitoylphosphatidylcholine or raft-like membranes were shown to induce increased ␣-helical structure in recombinant Syrian hamster PrP-(90 -231) at pH 7.0 (18, 19). Membrane interactio...

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“…For immunoprecipitation, untreated or PK-or DE-treated NHs and sCJDHs were centrifuged at 3000 ϫ g for 15 min at 4°C, and the supernatant was subjected to immunoprecipitation with either anti-ferritin or anti-PrP antibodies 6H4 or 8H4, as described previously (Mishra et al, 2002). The protein complexes were eluted from protein A beads with low pH glycine buffer, the pH was adjusted, and small aliquots of immunoprecipitated samples were frozen for additional use.…”

Section: Methodsmentioning

confidence: 99%