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Structural similarities between C6 and C7 of human complement.

E R Podack, W P Kolb, A F Esser

    Journal of Immunology (Baltimore, Md. : 1950)
    |September 1, 1979
    PubMed
    Summary
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    Researchers developed a novel affinity chromatography method to isolate human complement components C6 and C7. This technique yields pure, active proteins, suggesting a shared evolutionary origin for C6 and C7.

    Area of Science:

    • Biochemistry
    • Immunology
    • Proteomics

    Background:

    • Human complement system components C6 and C7 play crucial roles in immune responses.
    • Efficient isolation methods are essential for studying their structure and function.

    Purpose of the Study:

    • To develop and validate a new method for isolating human complement components C6 and C7.
    • To compare the biochemical and biophysical properties of isolated C6 and C7.

    Main Methods:

    • Affinity chromatography using anti-C6 and anti-C7 antibodies coupled to Sepharose.
    • Hemolytic activity assays.
    • Amino acid composition analysis.
    • Electrophoresis.
    • Isoelectric focusing.
    • UV spectroscopy.

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  • Circular dichroism (CD) spectroscopy.

    • Main Results:

    • A novel affinity chromatography method successfully isolated homogeneous, hemolytically active C6 and C7 in 25% yield.
    • Isolated C6 and C7 exhibit highly similar amino acid compositions and molecular weights (approx. 124,800 Da for C6, 120,800 Da for C7).
    • Both proteins are single-chain glycoproteins, migrating as beta 2-globulins, and display polymorphism via isoelectric focusing (pI 6.15-6.7).
    • Spectroscopic analyses (UV, CD) reveal minor differences in extinction coefficients and secondary structure content (C6: 8% alpha-helix, 10% beta-structure; C7: 10% alpha-helix, 14% beta-structure).

    Conclusions:

    • The developed affinity chromatography method provides an efficient means to obtain pure and active human C6 and C7.
    • The significant structural similarities between C6 and C7 support the hypothesis of their evolution from a common ancestral gene.