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Terminal components of carp complement constituting a membrane attack complex

M Nakao1, T Uemura, T Yano

  • 1Laboratory of Marine Biochemistry, Faculty of Agriculture, Kyushu University, Fukuoka, Japan.

Molecular Immunology
|August 1, 1996
PubMed
Summary
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The carp complement membrane attack complex (MAC) was purified and analyzed, revealing its composition and structure. This study demonstrates that bony fish complement utilizes a similar terminal pathway to mammals, involving components C5 through C9.

Area of Science:

  • Immunology
  • Biochemistry
  • Comparative Biology

Background:

  • The terminal pathway of the complement system forms the membrane attack complex (MAC), crucial for immune defense.
  • Understanding the MAC's structure and function in non-mammalian vertebrates provides insights into its evolutionary conservation.

Purpose of the Study:

  • To characterize the membrane attack complex (MAC) of carp (bony fish) complement.
  • To identify the individual protein components of carp MAC and determine their molar ratios.
  • To compare the carp MAC structure with that of mammals.

Main Methods:

  • Purification of carp MAC from rabbit erythrocytes lysed by carp serum using deoxycholate extraction and chromatography.
  • Analysis of carp MAC composition using two-dimensional SDS-PAGE.

Related Experiment Videos

  • Identification of protein subunits via Western blotting and N-terminal amino acid sequencing.
  • Densitometric scanning of gels to determine molar ratios of MAC components.

    • Main Results:

    • Carp MAC was resolved into eight distinct protein bands on two-dimensional SDS-PAGE.
    • Components C9, C8 (alpha, beta, gamma), C5b (alpha, beta), C6, and C7 were identified based on molecular weight, N-terminal sequences, and structural similarities to mammalian counterparts.
    • The molar ratio of C5b:C6:C7:C8:C9 in carp MAC was determined to be 1:1:1:1:4.

    Conclusions:

    • The carp MAC is composed of terminal complement components C5b through C9.
    • The structural and compositional analysis suggests that bony fish utilize a conserved cytolytic pathway similar to that of mammals.
    • This study provides evidence for the evolutionary conservation of the terminal complement pathway across vertebrates.