Detection of Myelin Basic Protein Isoforms by Organic Concentration
Identifieur interne : 001004 ( Main/Exploration ); précédent : 001003; suivant : 001005Detection of Myelin Basic Protein Isoforms by Organic Concentration
Auteurs : Jorma A. M Tt [Finlande] ; Eleanor T. Coffey [Finlande] ; Jorma A. Hermonen [Finlande] ; Aimo A. Salmi [Finlande] ; Ari E. Hinkkanen [Finlande]Source :
- Biochemical and Biophysical Research Communications [ 0006-291X ] ; 1997.
English descriptors
- KwdEn :
- CNS, central nervous system, EAE, EAE, experimental autoimmune encephalomyelitis, ECL, enhanced chemiluminescence, M, molecular mass, MBP, myelin basic protein, MS, multiple sclerosis, bMBP, bovine MBPchMBP, chicken MBP, fMBP, fish MBP, gpMBP, guinea-pig MBP, hMBP, human MBP, isoforms, mMBP, mouse MBP, myelin basic protein, organic extraction, purification, rMBP, ratMBP, rbMBP, rabbit MBP, sMBP, swine MBP.
Abstract
An effective technique was developed, which allowed rapid isolation of highly pure myelin basic protein (MBP) including its distinct isoforms. The procedure employs homogenization of central nervous system (CNS) tissue in chloroform, which specifically extracts MBP. Subsequently, methanol was used to convert the protein susceptible to quantitative transfer into the acidic aqueous phase. MBP was purified from bovine, chicken, fish, human, guinea-pig, mouse, rabbit, rat, and swine brains. Analysis on SDS-PAGE and immunoblotting using polyclonal MBP-specific serum recognized proteins corresponding to the sizes of previously identified MBP isoforms of 21.5, 18.5, 17.2, and 14.2 kDa and three predicted isoforms of 20.2, 16.0, and 13 kDa. The MBP obtained was readily soluble in water and possessed the capacity to induce experimental autoimmune encephalomyelitis in susceptible mice. The protein was also suitable for use as a substrate for protein kinases.
Url:
DOI: 10.1006/bbrc.1997.7318
Affiliations:
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Hermonen</name><affiliation wicri:level="4"><country xml:lang="fr">Finlande</country><wicri:regionArea>Department of Medical Biochemistry and Turku Centre for Biotechnology, University of Turku, Turku</wicri:regionArea><placeName><settlement type="city">Turku</settlement><region type="région" nuts="2">Finlande occidentale</region></placeName><orgName type="university">Université de Turku</orgName></affiliation></author><author><name sortKey="Salmi, Aimo A" sort="Salmi, Aimo A" uniqKey="Salmi A" first="Aimo A." last="Salmi">Aimo A. 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The procedure employs homogenization of central nervous system (CNS) tissue in chloroform, which specifically extracts MBP. Subsequently, methanol was used to convert the protein susceptible to quantitative transfer into the acidic aqueous phase. MBP was purified from bovine, chicken, fish, human, guinea-pig, mouse, rabbit, rat, and swine brains. Analysis on SDS-PAGE and immunoblotting using polyclonal MBP-specific serum recognized proteins corresponding to the sizes of previously identified MBP isoforms of 21.5, 18.5, 17.2, and 14.2 kDa and three predicted isoforms of 20.2, 16.0, and 13 kDa. The MBP obtained was readily soluble in water and possessed the capacity to induce experimental autoimmune encephalomyelitis in susceptible mice. The protein was also suitable for use as a substrate for protein kinases.</div></front></TEI><affiliations><list><country><li>Finlande</li></country><region><li>Finlande occidentale</li></region><settlement><li>Turku</li></settlement><orgName><li>Université de Turku</li></orgName></list><tree><country name="Finlande"><region name="Finlande occidentale"><name sortKey="M Tt, Jorma A" sort="M Tt, Jorma A" uniqKey="M Tt J" first="Jorma A." last="M Tt">Jorma A. M Tt</name></region><name sortKey="Coffey, Eleanor T" sort="Coffey, Eleanor T" uniqKey="Coffey E" first="Eleanor T." last="Coffey">Eleanor T. Coffey</name><name sortKey="Hermonen, Jorma A" sort="Hermonen, Jorma A" uniqKey="Hermonen J" first="Jorma A." last="Hermonen">Jorma A. Hermonen</name><name sortKey="Hinkkanen, Ari E" sort="Hinkkanen, Ari E" uniqKey="Hinkkanen A" first="Ari E." last="Hinkkanen">Ari E. Hinkkanen</name><name sortKey="Salmi, Aimo A" sort="Salmi, Aimo A" uniqKey="Salmi A" first="Aimo A." last="Salmi">Aimo A. 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