Strategies for the qualitative and quantitative analysis of metallothionein isoforms by capillary electrophoresis

J H Beattie

    Research output: Contribution to journalArticle

    38 Citations (Scopus)

    Abstract

    Metallothioneins (MT) are a heterogeneous family of proteins coded from multiple linked genes whose individual expression can be readily determined at the transcriptional level by assay of mRNA using specific DNA probes targeted against the untranslated regions. Analysis of translated protein products is more challenging since sequence variation, between MT isoforms can be as little as one in 61 amino acids. Separation of these isoforms therefore requires a high resolution technique which can exploit minor differences in charge or hydrophobicity. The use of capillary electrophoresis (CE) for MT isoform separation is reviewed, including practical information on capillaries and electrolyte conditions which are most suitable for qualitative evaluation of purified MT and also for quantification of isoforms from biological tissues. Methods of sample preparation are discussed, as are methods of characterising the separated components and enhancing sensitivity of their detection. (C) 1998 Elsevier Science B.V. All rights reserved.

    Original languageEnglish
    Pages (from-to)255-270
    Number of pages16
    JournalTalanta
    Volume46
    Issue number2
    Publication statusPublished - Jun 1998

    Keywords

    • metallothionein
    • isoform separation
    • capillary electrophoresis
    • MECC
    • SPE-CE
    • MALDI-TOFMS
    • PERFORMANCE LIQUID-CHROMATOGRAPHY
    • ZONE ELECTROPHORESIS
    • ONLINE PRECONCENTRATION
    • MASS-SPECTROMETRY
    • SEPARATION
    • PURIFICATION
    • PROTEIN
    • LOCUS
    • BRAIN

    Cite this

    Strategies for the qualitative and quantitative analysis of metallothionein isoforms by capillary electrophoresis. / Beattie, J H .

    In: Talanta, Vol. 46, No. 2, 06.1998, p. 255-270.

    Research output: Contribution to journalArticle

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    abstract = "Metallothioneins (MT) are a heterogeneous family of proteins coded from multiple linked genes whose individual expression can be readily determined at the transcriptional level by assay of mRNA using specific DNA probes targeted against the untranslated regions. Analysis of translated protein products is more challenging since sequence variation, between MT isoforms can be as little as one in 61 amino acids. Separation of these isoforms therefore requires a high resolution technique which can exploit minor differences in charge or hydrophobicity. The use of capillary electrophoresis (CE) for MT isoform separation is reviewed, including practical information on capillaries and electrolyte conditions which are most suitable for qualitative evaluation of purified MT and also for quantification of isoforms from biological tissues. Methods of sample preparation are discussed, as are methods of characterising the separated components and enhancing sensitivity of their detection. (C) 1998 Elsevier Science B.V. All rights reserved.",
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