DOI: 10.1002/pro.5560011014. Purification and properties of the cellular prion protein from Syrian hamster brain

Purification and properties of the cellular prion protein from Syrian hamster brain

10.1002/pro.5560011014

Crossref journal-article

Wiley

Protein Science (311)

Abstract

AbstractThe cellular prion protein (PrPC) is encoded by a chromosomal gene, and its scrapie isoform (PrPSc) features in all aspects of the prion diseases. Prior to the studies reported here, purification of PrPC has only been accomplished using immunoaffinity chromatography yielding small amounts of protein. Brain homogenates contain two PrPC forms designated PrPC‐I and ‐II. These proteins were purified from a microsomal fraction by detergent extraction and separated by immobilized Cu2+ ion affinity chromatography. PrPC‐II appears to be generated from PrPC‐I by limited proteolysis of the N‐terminus. Fractions enriched for PrPC‐I were purified further by cation‐exchange chromatography and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE). Greater than 90% of the final product migrated as a broad band of Mr 33–35 kDa as judged by silver staining after SDS‐PAGE. Digestion of PrPC‐I with peptide‐N‐glycosidase (PNGase) compressed the band and shifted its mobility giving an Mr of 27 kDa. The protocol described should be amenable to large‐scale preparation of PrPC, enabling physical comparisons of PrPC and PrPSc.

Bibliography

Pan, K., Stahl, N., & Prusiner, S. B. (1992). Purification and properties of the cellular prion protein from Syrian hamster brain. Protein Science, 1(10), 1343â1352. Portico.

Authors 3

Keh‐Ming Pan (first)
Neil Stahl (additional)
Stanley B. Prusiner (additional)

References 60 Referenced 150

10.1016/0003-2697(84)90347-6
{'key': 'e_1_2_1_3_1', 'first-page': '3624', 'article-title': 'Facile resolution of alpha‐fetoproteins and serum albumins by immobilized metal affinity chromatography', 'volume': '47', 'author': 'Andersson L.', 'year': '1987', 'journal-title': 'Cancer Res.'} / Cancer Res. / Facile resolution of alpha‐fetoproteins and serum albumins by immobilized metal affinity chromatography by Andersson L. (1987)
10.1016/0003-2697(90)90405-X
10.1093/infdis/154.3.518
10.4049/jimmunol.140.4.1188 / J. Immunol. / Characterization of prion proteins with monospecific antisera to synthetic peptides by Barry R.A. (1988)
10.1016/0092-8674(86)90662-8
10.1038/325581a0
10.1093/infdis/158.6.1198
10.1016/0003-9861(87)90380-8
10.1128/jvi.53.2.596-606.1985 / J. Virol. / Scrapie PrP 27–30 is a sialoglycoprotein by Bolton D.C. (1985)
10.1128/jvi.65.7.3667-3675.1991
10.1083/jcb.110.3.743
10.1016/0003-2697(76)90527-3
10.1042/bj1950015
10.1016/S0021-9258(18)55257-1
10.1021/bi00245a003
10.1016/S0140-6736(61)92008-6
10.1038/315331a0
10.1021/bi00447a017
10.1073/pnas.84.12.4017
Gasset M. Baldwin M.A. Fletterick R.J. &Prusiner S.B.(1992).Perturbation of the secondary structure of the scrapie prion protein under conditions associated with changes in infectivity.Proc. Natl. Acad. Sci. in press. (10.1073/pnas.90.1.1)
10.1073/pnas.87.7.2476
10.1099/0022-1317-72-1-201
10.1016/0003-9861(89)90409-8
10.1073/pnas.88.17.7664
10.1073/pnas.86.6.1811
10.1002/j.1460-2075.1986.tb04539.x
10.1111/j.1432-1033.1988.tb13883.x
10.1038/338342a0
10.1128/jvi.61.12.3688-3693.1987
{'key': 'e_1_2_1_32_1', 'first-page': '1', 'article-title': 'Scrapie prion proteins are synthesized in neurons', 'volume': '122', 'author': 'Kretzschmar H.A.', 'year': '1986', 'journal-title': 'Am. J. Pathol.'} / Am. J. Pathol. / Scrapie prion proteins are synthesized in neurons by Kretzschmar H.A. (1986)
10.1038/227680a0
10.1128/MCB.10.3.1153
{'key': 'e_1_2_1_35_1', 'first-page': '622', 'article-title': 'Ultrastructural localization of scrapie prion proteins in cytoplasmic vesicles of infected cultured cells', 'volume': '65', 'author': 'McKinley M.P.', 'year': '1991', 'journal-title': 'Lab. Invest.'} / Lab. Invest. / Ultrastructural localization of scrapie prion proteins in cytoplasmic vesicles of infected cultured cells by McKinley M.P. (1991)
10.1016/0003-2697(81)90136-6
10.1073/pnas.83.8.2310
10.1073/pnas.85.24.9811
10.1016/0092-8674(85)90333-2
10.1038/258598a0
10.1126/science.1675487
10.1021/bi00269a050
10.1016/0092-8674(84)90533-6
10.1021/bi00562a028
10.1021/bi00616a020
10.1021/bi00616a021
10.1016/0092-8674(83)90168-X
10.1016/0092-8674(90)90134-Z
{'key': 'e_1_2_1_49_1', 'first-page': '320', 'article-title': 'Genomic structure of the human prion protein gene', 'volume': '49', 'author': 'Puckett C.', 'year': '1991', 'journal-title': 'Am. J. Hum. Genet.'} / Am. J. Hum. Genet. / Genomic structure of the human prion protein gene by Puckett C. (1991)
10.4049/jimmunol.147.10.3568 / J. Immunol. / Epitope mapping of the Syrian hamster prion protein utilizing chimeric and mutant genes in a vaccinia virus expression system by Rogers M. (1991)
10.1073/pnas.87.16.6373
10.1021/bi00136a600
{'key': 'e_1_2_1_53_1', 'volume-title': 'Prion Diseases of Humans and Animals', 'author': 'Stahl N.', 'year': '1992'} / Prion Diseases of Humans and Animals by Stahl N. (1992)
10.1016/0092-8674(87)90150-4
10.1016/0167-7799(85)90068-X
10.1002/bies.950100508
10.1083/jcb.110.6.2117
10.1073/pnas.76.9.4350
10.1111/j.1432-1033.1988.tb14246.x
10.1016/0092-8674(87)90134-6
10.1126/science.6177036

Dates

Type	When
Created	16 years, 6 months ago (Feb. 9, 2009, 6:08 p.m.)
Deposited	1 year, 10 months ago (Sept. 27, 2023, 12:12 p.m.)
Indexed	3 months ago (May 21, 2025, 3:31 a.m.)
Issued	32 years, 10 months ago (Oct. 1, 1992)
Published	32 years, 10 months ago (Oct. 1, 1992)
Published Online	16 years, 7 months ago (Dec. 31, 2008)
Published Print	32 years, 10 months ago (Oct. 1, 1992)

Funders 0

None

BibTeX

@article{Pan_1992, title={Purification and properties of the cellular prion protein from Syrian hamster brain}, volume={1}, ISSN={1469-896X}, url={http://dx.doi.org/10.1002/pro.5560011014}, DOI={10.1002/pro.5560011014}, number={10}, journal={Protein Science}, publisher={Wiley}, author={Pan, Keh‐Ming and Stahl, Neil and Prusiner, Stanley B.}, year={1992}, month=oct, pages={1343–1352} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        5,
        21
      ]
    ],
    "date-time": "2025-05-21T07:31:09Z",
    "timestamp": 1747812669328
  },
  "reference-count": 60,
  "publisher": "Wiley",
  "issue": "10",
  "license": [
    {
      "start": {
        "date-parts": [
          [
            2008,
            12,
            31
          ]
        ],
        "date-time": "2008-12-31T00:00:00Z",
        "timestamp": 1230681600000
      },
      "content-version": "vor",
      "delay-in-days": 5935,
      "URL": "http://onlinelibrary.wiley.com/termsAndConditions#vor"
    }
  ],
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        1992,
        10
      ]
    ]
  },
  "abstract": "<jats:title>Abstract</jats:title><jats:p>The cellular prion protein (PrP<jats:sup><jats:italic>C</jats:italic></jats:sup>) is encoded by a chromosomal gene, and its scrapie isoform (PrP<jats:sup>Sc</jats:sup>) features in all aspects of the prion diseases. Prior to the studies reported here, purification of PrP<jats:sup><jats:italic>C</jats:italic></jats:sup> has only been accomplished using immunoaffinity chromatography yielding small amounts of protein. Brain homogenates contain two PrP<jats:sup><jats:italic>C</jats:italic></jats:sup> forms designated PrP<jats:sup><jats:italic>C</jats:italic></jats:sup>\u2010I and \u2010II. These proteins were purified from a microsomal fraction by detergent extraction and separated by immobilized Cu<jats:sup>2+</jats:sup> ion affinity chromatography. PrP<jats:sup><jats:italic>C</jats:italic></jats:sup>\u2010II appears to be generated from PrP<jats:sup><jats:italic>C</jats:italic></jats:sup>\u2010I by limited proteolysis of the N\u2010terminus. Fractions enriched for PrP<jats:sup><jats:italic>C</jats:italic></jats:sup>\u2010I were purified further by cation\u2010exchange chromatography and sodium dodecyl sulfate\u2010polyacrylamide gel electrophoresis (SDS\u2010PAGE). Greater than 90% of the final product migrated as a broad band of <jats:italic>M<jats:sub>r</jats:sub></jats:italic> 33\u201335 kDa as judged by silver staining after SDS\u2010PAGE. Digestion of PrP<jats:sup><jats:italic>C</jats:italic></jats:sup>\u2010I with peptide\u2010<jats:italic>N</jats:italic>\u2010glycosidase (PNGase) compressed the band and shifted its mobility giving an <jats:italic>M<jats:sub>r</jats:sub></jats:italic> of 27 kDa. The protocol described should be amenable to large\u2010scale preparation of PrP<jats:sup><jats:italic>C</jats:italic></jats:sup>, enabling physical comparisons of PrP<jats:sup><jats:italic>C</jats:italic></jats:sup> and PrP<jats:sup>Sc</jats:sup>.</jats:p>",
  "DOI": "10.1002/pro.5560011014",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2009,
        2,
        9
      ]
    ],
    "date-time": "2009-02-09T23:08:43Z",
    "timestamp": 1234220923000
  },
  "page": "1343-1352",
  "source": "Crossref",
  "is-referenced-by-count": 150,
  "title": "Purification and properties of the cellular prion protein from Syrian hamster brain",
  "prefix": "10.1002",
  "volume": "1",
  "author": [
    {
      "given": "Keh\u2010Ming",
      "family": "Pan",
      "sequence": "first",
      "affiliation": []
    },
    {
      "given": "Neil",
      "family": "Stahl",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Stanley B.",
      "family": "Prusiner",
      "sequence": "additional",
      "affiliation": []
    }
  ],
  "member": "311",
  "published-online": {
    "date-parts": [
      [
        2008,
        12,
        31
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_2_1_2_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0003-2697(84)90347-6"
    },
    {
      "key": "e_1_2_1_3_1",
      "first-page": "3624",
      "article-title": "Facile resolution of alpha\u2010fetoproteins and serum albumins by immobilized metal affinity chromatography",
      "volume": "47",
      "author": "Andersson L.",
      "year": "1987",
      "journal-title": "Cancer Res."
    },
    {
      "key": "e_1_2_1_4_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0003-2697(90)90405-X"
    },
    {
      "key": "e_1_2_1_5_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/infdis/154.3.518"
    },
    {
      "key": "e_1_2_1_6_1",
      "doi-asserted-by": "crossref",
      "first-page": "1188",
      "DOI": "10.4049/jimmunol.140.4.1188",
      "article-title": "Characterization of prion proteins with monospecific antisera to synthetic peptides",
      "volume": "140",
      "author": "Barry R.A.",
      "year": "1988",
      "journal-title": "J. Immunol."
    },
    {
      "key": "e_1_2_1_7_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(86)90662-8"
    },
    {
      "key": "e_1_2_1_8_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/325581a0"
    },
    {
      "key": "e_1_2_1_9_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/infdis/158.6.1198"
    },
    {
      "key": "e_1_2_1_10_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0003-9861(87)90380-8"
    },
    {
      "key": "e_1_2_1_11_1",
      "doi-asserted-by": "crossref",
      "first-page": "596",
      "DOI": "10.1128/jvi.53.2.596-606.1985",
      "article-title": "Scrapie PrP 27\u201330 is a sialoglycoprotein",
      "volume": "53",
      "author": "Bolton D.C.",
      "year": "1985",
      "journal-title": "J. Virol."
    },
    {
      "key": "e_1_2_1_12_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.65.7.3667-3675.1991"
    },
    {
      "key": "e_1_2_1_13_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1083/jcb.110.3.743"
    },
    {
      "key": "e_1_2_1_14_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0003-2697(76)90527-3"
    },
    {
      "key": "e_1_2_1_15_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1042/bj1950015"
    },
    {
      "key": "e_1_2_1_16_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9258(18)55257-1"
    },
    {
      "key": "e_1_2_1_17_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/bi00245a003"
    },
    {
      "key": "e_1_2_1_18_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0140-6736(61)92008-6"
    },
    {
      "key": "e_1_2_1_19_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/315331a0"
    },
    {
      "key": "e_1_2_1_20_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/bi00447a017"
    },
    {
      "key": "e_1_2_1_21_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.84.12.4017"
    },
    {
      "key": "e_1_2_1_22_1",
      "doi-asserted-by": "crossref",
      "unstructured": "Gasset M. Baldwin M.A. Fletterick R.J. &Prusiner S.B.(1992).Perturbation of the secondary structure of the scrapie prion protein under conditions associated with changes in infectivity.Proc. Natl. Acad. Sci. in press.",
      "DOI": "10.1073/pnas.90.1.1"
    },
    {
      "key": "e_1_2_1_23_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.87.7.2476"
    },
    {
      "key": "e_1_2_1_24_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1099/0022-1317-72-1-201"
    },
    {
      "key": "e_1_2_1_25_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0003-9861(89)90409-8"
    },
    {
      "key": "e_1_2_1_26_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.88.17.7664"
    },
    {
      "key": "e_1_2_1_27_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.86.6.1811"
    },
    {
      "key": "e_1_2_1_28_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/j.1460-2075.1986.tb04539.x"
    },
    {
      "key": "e_1_2_1_29_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1432-1033.1988.tb13883.x"
    },
    {
      "key": "e_1_2_1_30_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/338342a0"
    },
    {
      "key": "e_1_2_1_31_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.61.12.3688-3693.1987"
    },
    {
      "key": "e_1_2_1_32_1",
      "first-page": "1",
      "article-title": "Scrapie prion proteins are synthesized in neurons",
      "volume": "122",
      "author": "Kretzschmar H.A.",
      "year": "1986",
      "journal-title": "Am. J. Pathol."
    },
    {
      "key": "e_1_2_1_33_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/227680a0"
    },
    {
      "key": "e_1_2_1_34_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/MCB.10.3.1153"
    },
    {
      "key": "e_1_2_1_35_1",
      "first-page": "622",
      "article-title": "Ultrastructural localization of scrapie prion proteins in cytoplasmic vesicles of infected cultured cells",
      "volume": "65",
      "author": "McKinley M.P.",
      "year": "1991",
      "journal-title": "Lab. Invest."
    },
    {
      "key": "e_1_2_1_36_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0003-2697(81)90136-6"
    },
    {
      "key": "e_1_2_1_37_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.83.8.2310"
    },
    {
      "key": "e_1_2_1_38_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.85.24.9811"
    },
    {
      "key": "e_1_2_1_39_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(85)90333-2"
    },
    {
      "key": "e_1_2_1_40_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/258598a0"
    },
    {
      "key": "e_1_2_1_41_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.1675487"
    },
    {
      "key": "e_1_2_1_42_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/bi00269a050"
    },
    {
      "key": "e_1_2_1_43_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(84)90533-6"
    },
    {
      "key": "e_1_2_1_44_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/bi00562a028"
    },
    {
      "key": "e_1_2_1_45_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/bi00616a020"
    },
    {
      "key": "e_1_2_1_46_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/bi00616a021"
    },
    {
      "key": "e_1_2_1_47_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(83)90168-X"
    },
    {
      "key": "e_1_2_1_48_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(90)90134-Z"
    },
    {
      "key": "e_1_2_1_49_1",
      "first-page": "320",
      "article-title": "Genomic structure of the human prion protein gene",
      "volume": "49",
      "author": "Puckett C.",
      "year": "1991",
      "journal-title": "Am. J. Hum. Genet."
    },
    {
      "key": "e_1_2_1_50_1",
      "doi-asserted-by": "crossref",
      "first-page": "3568",
      "DOI": "10.4049/jimmunol.147.10.3568",
      "article-title": "Epitope mapping of the Syrian hamster prion protein utilizing chimeric and mutant genes in a vaccinia virus expression system",
      "volume": "147",
      "author": "Rogers M.",
      "year": "1991",
      "journal-title": "J. Immunol."
    },
    {
      "key": "e_1_2_1_51_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.87.16.6373"
    },
    {
      "key": "e_1_2_1_52_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/bi00136a600"
    },
    {
      "key": "e_1_2_1_53_1",
      "volume-title": "Prion Diseases of Humans and Animals",
      "author": "Stahl N.",
      "year": "1992"
    },
    {
      "key": "e_1_2_1_54_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(87)90150-4"
    },
    {
      "key": "e_1_2_1_55_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0167-7799(85)90068-X"
    },
    {
      "key": "e_1_2_1_56_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/bies.950100508"
    },
    {
      "key": "e_1_2_1_57_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1083/jcb.110.6.2117"
    },
    {
      "key": "e_1_2_1_58_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.76.9.4350"
    },
    {
      "key": "e_1_2_1_59_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1432-1033.1988.tb14246.x"
    },
    {
      "key": "e_1_2_1_60_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(87)90134-6"
    },
    {
      "key": "e_1_2_1_61_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.6177036"
    }
  ],
  "container-title": "Protein Science",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fpro.5560011014",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "text-mining"
    },
    {
      "URL": "https://onlinelibrary.wiley.com/doi/pdf/10.1002/pro.5560011014",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2023,
        9,
        27
      ]
    ],
    "date-time": "2023-09-27T16:12:46Z",
    "timestamp": 1695831166000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://onlinelibrary.wiley.com/doi/10.1002/pro.5560011014"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        1992,
        10
      ]
    ]
  },
  "references-count": 60,
  "journal-issue": {
    "issue": "10",
    "published-print": {
      "date-parts": [
        [
          1992,
          10
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1002/pro.5560011014"
  ],
  "URL": "http://dx.doi.org/10.1002/pro.5560011014",
  "relation": {},
  "ISSN": [
    "0961-8368",
    "1469-896X"
  ],
  "subject": [],
  "container-title-short": "Protein Science",
  "published": {
    "date-parts": [
      [
        1992,
        10
      ]
    ]
  }
}