Insoluble Aggregates and Protease-resistant Conformers of Prion Protein in Uninfected Human Brains

Yuan, Jing; Xiao, Xinxin; McGeehan, J.E.; Dong, Zhiqian; Calì, Ignazio; Fujioka, Hisashi; Kong, Qingzhong; Kneale, Geoff; Gambetti, Pierluigi; Zou, Wen-Quan

doi:10.1074/jbc.m602238200

Cited by 117 publications

(189 citation statements)

References 62 publications

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“…In this study, we also found that the previously described 18-kDa C1 PrP C fragment (44) shows a relatively high degree of protease resistance in white matter control samples. The observation that the C1 fragment has a higher PK resistance when compared with full-length PrP C has been already underlined in previous studies (52)(53)(54). We extended these observations and found that in white matter samples, the fragment is still detectable after PK digestion at an enzyme concentration of 20 units/ml, which is associated with a complete degradation of full-length PrP C .…”

Section: Discussionsupporting

confidence: 80%

Characterization of Truncated Forms of Abnormal Prion Protein in Creutzfeldt-Jakob Disease

Notari

Strammiello

Capellari

et al. 2008

Journal of Biological Chemistry

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In prion disease, the abnormal conformer of the cellular prion protein, PrP Sc , deposits in fibrillar protein aggregates in brain and other organs. Limited exposure of PrP Sc to proteolytic digestion in vitro generates a core fragment of 19 -21 kDa, named PrP27-30, which is also found in vivo. Recent evidence indicates that abnormal truncated fragments other than PrP27-30 may form in prion disease either in vivo or in vitro. We characterized a novel protease-resistant PrP fragment migrating 2-3 kDa faster than PrP27-30 in Creutzfeldt-Jakob disease (CJD) brains. The fragment has a size of about 18.5 kDa when associated with PrP27-30 type 1 (21 kDa) and of 17 kDa when associated with type 2 (19 kDa). Molecular mass and epitope mapping showed that the two fragments share the primary N-terminal sequence with PrP27-30 types 1 and 2, respectively, but lack a few amino acids at the very end of C terminus together with the glycosylphosphatidylinositol anchor. The amounts of the 18.5-or 17-kDa fragments and the previously described 13-kDa PrP Sc C-terminal fragment relatively to the PrP27-30 signal significantly differed among CJD subtypes.

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Section: Discussionsupporting

confidence: 80%

Characterization of Truncated Forms of Abnormal Prion Protein in Creutzfeldt-Jakob Disease

Notari

Strammiello

Capellari

et al. 2008

Journal of Biological Chemistry

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“…G5p is capable of capturing both rPrP Sc and PK-sensitive PrP Sc (sPrP Sc ) species. 14,20,21 Although the g5p beads precipitated the abnormal PrP Sc from 5 l of CJD brain homogenate, PrP was not detectable in samples captured by g5p from 6 ml of amniotic fluid and 6 ml of uterine and placental homogenates each in both CJD and non-CJD controls ( Figure 3B and data not shown). PrP was immunoprecipitable by 6H4-conjugated beads ( Figure 3B).…”

Section: Detection Of Prp In the Uterus Placenta And Amniotic Fluidmentioning

confidence: 90%

“…Anti-PrP antibodies, including rabbit anti-C-terminal antiserum immunoreactive to human PrP residues 220 to 231, mouse monoclonal antibody 3F4 against human PrP residues 109 to 112, mouse monoclonal antibody 1E4 against human PrP 97 to 108 (Cell Sciences, Canton, MA), and mouse monoclonal antibody 6H4 against human PrP 145 to 152 (Prionics AG, Zurich, Switzerland) were used. [11][12][13][14] Human Tissues Consent to use autopsy and biopsy material for research purposes was obtained for all samples. The neuropathological examination of biopsy and autopsy specimens of the proband was performed at the Indiana Alzheimer Disease Center, Indianapolis, IN, and at the National …”

Section: Reagents and Antibodiesmentioning

confidence: 99%

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Failure to Detect the Presence of Prions in the Uterine and Gestational Tissues from a Gravida with Creutzfeldt-Jakob Disease

Xiao

Miravalle

Yuan

et al. 2009

The American Journal of Pathology

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The vertical transmission of a prion disease from infected mothers to their offspring is believed to be one of the routes for the natural spread of animal prion diseases. Supporting this notion is the observation that prion infectivity occurs in the placenta of infected ewes. Furthermore, the prion protein (PrP), both in its cellular form (PrP C ) and its pathological isoform (PrP Sc ), has been observed at the fetal-maternal interface of scrapie-infected sheep. However, whether these features of prion infectivity also hold true for human prion diseases is currently unknown. To begin to address such an important question, we examined PrP in the uterus as well as gestational tissues, including the placenta and amniotic fluid, in a pregnant woman with sporadic Creutzfeldt-Jakob disease (CJD). Although the proteinase K (PK)-resistant prion protein, PrP27-30, was present in the brain tissues of the mother, the PrP detected in the uterus, placenta, and amniotic fluid was sensitive to PK digestion. Unlike PrP C in the brain and adjacent cerebrospinal fluid, the predominant PrP species in the reproductive and gestational tissues were N-terminally truncated, similar to urine PrP. Our study did not detect abnormal PrP in the reproductive and gestational tissues in this case of CJD. Nevertheless, examination by a highly sensitive bioassay is ongoing to ascertain possible prion infectivity from CJD in the amniotic fluid.

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“…The types of protein that aggregate with the PrPreslike molecules are not known. Some plasma proteins are known to associate with the PrP, but it is possible that the PrP molecules in hamster plasma may also be a candidate for these plasma proteins (11,31,34). PK- resistant PrP molecules have recently been reported in uninfected human brains as well as in uninfected mouse and hamster brains and have been labeled a silent prion.…”

Section: Discussionmentioning

confidence: 99%

A Potential Blood Test for Transmissible Spongiform Encephalopathies by Detecting Carbohydrate‐Dependent Aggregates of PrPres‐Like Proteins in Scrapie‐Infected Hamster Plasma

Tsukui

Takata

Tadokoro

2007

Microbiology and Immunology

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PrPres has rarely been detected in blood (except in leukocytes) even in diseased animal models that are known to contain a large amount of PrPres in infected tissues. It seems likely that PrPres detection in blood is difficult because of the low titer of infectious material within the blood. Here, we demonstrate the detection of proteinase K‐resistant 3F4‐reactive protein in the plasma of scrapie‐infected hamsters but not in the plasma of mock‐infected hamsters by partial purification using a novel method termed “acidic SDS precipitation,” in conjunction with a highly sensitive chemiluminescence detection system used to show the presence of PrP at a concentration equivalent to 1.4 ×10–9 g of brain homogenate or 1.5 × 10–12 g (6.5 ×10–17 mol) of rPrP by conventional Western blotting. The 3F4‐reactive proteins in scrapie‐infected hamster plasma often resulted in multiple Mw protein bands occurring at higher Mw positions than the position of the di‐glycosyl PrP molecule. Mixing scrapie‐infected hamster brain homogenate with mock‐infected hamster plasma resulted in the formation of similar Mw positions for multiple 3F4‐reactive proteins. Predigestion of carbohydrate side chains from the proteins in the plasma or brain homogenate before mixing resulted in failure to obtain these multiple 3F4‐reactive proteins. These observations indicate that PrPres aggregated with other proteins in the plasma through carbohydrate side chains and was successfully detected in the plasma of scrapie‐infected hamsters. Counterparts in these aggregates with PrPres‐like proteins in scHaPl are not known but any that exist should resist the PK digestion.

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Insoluble Aggregates and Protease-resistant Conformers of Prion Protein in Uninfected Human Brains

Cited by 117 publications

References 62 publications

Characterization of Truncated Forms of Abnormal Prion Protein in Creutzfeldt-Jakob Disease

Characterization of Truncated Forms of Abnormal Prion Protein in Creutzfeldt-Jakob Disease

Failure to Detect the Presence of Prions in the Uterine and Gestational Tissues from a Gravida with Creutzfeldt-Jakob Disease

A Potential Blood Test for Transmissible Spongiform Encephalopathies by Detecting Carbohydrate‐Dependent Aggregates of PrPres‐Like Proteins in Scrapie‐Infected Hamster Plasma

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