Isolation of high-specific-activity subunits of cholera toxin by reversed-phase high-performance liquid chromatography

Stephen D. Pearson, James D. Dixon, Steven F. Northwehr, Alexander Kurosky

    Research output: Contribution to journalArticle

    4 Citations (Scopus)

    Abstract

    Facile, rapid procedures for the separation of active cholera toxin subunits were developed, based on high-performance liquid chromatography (HPLC) with a Nucleosil C8 reversed-phase column. These procedures were capable of completely resolving subunits A and B as well as S-carboxymethylated or reduced α-, γ-, and β-chains. The binding of HPLC-purified B subunit to GM1 ganglioside was essentially identical to that of cholera toxin when compared on a molar basis. The adenosine 5′-diphosphate-ribosyltransferase activity of HPLC-purified A subunit, reduced α-chain, or carboxymethyl α-chain was also determined to be reasonably high compared to that of cholera toxin or commercially prepared A subunit.

    Original languageEnglish (US)
    Pages (from-to)413-421
    Number of pages9
    JournalJournal of Chromatography A
    Volume359
    Issue numberC
    DOIs
    StatePublished - 1986

    Fingerprint

    Cholera Toxin
    High performance liquid chromatography
    Reverse-Phase Chromatography
    High Pressure Liquid Chromatography
    G(M1) Ganglioside
    Diphosphates
    Adenosine
    Adenosine Diphosphate

    ASJC Scopus subject areas

    • Analytical Chemistry
    • Clinical Biochemistry
    • Molecular Medicine

    Cite this

    Isolation of high-specific-activity subunits of cholera toxin by reversed-phase high-performance liquid chromatography. / Pearson, Stephen D.; Dixon, James D.; Northwehr, Steven F.; Kurosky, Alexander.

    In: Journal of Chromatography A, Vol. 359, No. C, 1986, p. 413-421.

    Research output: Contribution to journalArticle

    Pearson, Stephen D. ; Dixon, James D. ; Northwehr, Steven F. ; Kurosky, Alexander. / Isolation of high-specific-activity subunits of cholera toxin by reversed-phase high-performance liquid chromatography. In: Journal of Chromatography A. 1986 ; Vol. 359, No. C. pp. 413-421.
    @article{b7a1c561f77249e795bdcd488b753d23,
    title = "Isolation of high-specific-activity subunits of cholera toxin by reversed-phase high-performance liquid chromatography",
    abstract = "Facile, rapid procedures for the separation of active cholera toxin subunits were developed, based on high-performance liquid chromatography (HPLC) with a Nucleosil C8 reversed-phase column. These procedures were capable of completely resolving subunits A and B as well as S-carboxymethylated or reduced α-, γ-, and β-chains. The binding of HPLC-purified B subunit to GM1 ganglioside was essentially identical to that of cholera toxin when compared on a molar basis. The adenosine 5′-diphosphate-ribosyltransferase activity of HPLC-purified A subunit, reduced α-chain, or carboxymethyl α-chain was also determined to be reasonably high compared to that of cholera toxin or commercially prepared A subunit.",
    author = "Pearson, {Stephen D.} and Dixon, {James D.} and Northwehr, {Steven F.} and Alexander Kurosky",
    year = "1986",
    doi = "10.1016/0021-9673(86)80095-4",
    language = "English (US)",
    volume = "359",
    pages = "413--421",
    journal = "Journal of Chromatography",
    issn = "0021-9673",
    publisher = "Elsevier",
    number = "C",

    }

    TY - JOUR

    T1 - Isolation of high-specific-activity subunits of cholera toxin by reversed-phase high-performance liquid chromatography

    AU - Pearson, Stephen D.

    AU - Dixon, James D.

    AU - Northwehr, Steven F.

    AU - Kurosky, Alexander

    PY - 1986

    Y1 - 1986

    N2 - Facile, rapid procedures for the separation of active cholera toxin subunits were developed, based on high-performance liquid chromatography (HPLC) with a Nucleosil C8 reversed-phase column. These procedures were capable of completely resolving subunits A and B as well as S-carboxymethylated or reduced α-, γ-, and β-chains. The binding of HPLC-purified B subunit to GM1 ganglioside was essentially identical to that of cholera toxin when compared on a molar basis. The adenosine 5′-diphosphate-ribosyltransferase activity of HPLC-purified A subunit, reduced α-chain, or carboxymethyl α-chain was also determined to be reasonably high compared to that of cholera toxin or commercially prepared A subunit.

    AB - Facile, rapid procedures for the separation of active cholera toxin subunits were developed, based on high-performance liquid chromatography (HPLC) with a Nucleosil C8 reversed-phase column. These procedures were capable of completely resolving subunits A and B as well as S-carboxymethylated or reduced α-, γ-, and β-chains. The binding of HPLC-purified B subunit to GM1 ganglioside was essentially identical to that of cholera toxin when compared on a molar basis. The adenosine 5′-diphosphate-ribosyltransferase activity of HPLC-purified A subunit, reduced α-chain, or carboxymethyl α-chain was also determined to be reasonably high compared to that of cholera toxin or commercially prepared A subunit.

    UR - http://www.scopus.com/inward/record.url?scp=0022556355&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=0022556355&partnerID=8YFLogxK

    U2 - 10.1016/0021-9673(86)80095-4

    DO - 10.1016/0021-9673(86)80095-4

    M3 - Article

    C2 - 3016001

    AN - SCOPUS:0022556355

    VL - 359

    SP - 413

    EP - 421

    JO - Journal of Chromatography

    JF - Journal of Chromatography

    SN - 0021-9673

    IS - C

    ER -