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Structural basis for the C4d.1/C4d.2 serologic allotypes of murine complement component C4.

P A Taillon-Miller1, D C Shreffler

  • 1Department of Genetics, Washington University School of Medicine, St. Louis, MO 63110.

Journal of Immunology (Baltimore, Md. : 1950)
|October 1, 1988
PubMed
Summary

The C4d.1 and C4d.2 specificities, defined by H-2 alloantigens, differ due to a single amino acid substitution. This glutamine-to-arginine change in the C4d fragment explains their distinct serological properties.

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Area of Science:

  • Immunogenetics
  • Molecular immunology
  • Complement system

Background:

  • C4d.1 and C4d.2 are H-2 associated alloantigens defined serologically.
  • These specificities represent allotypes of complement component C4, with the determinant on the C4d fragment.
  • Previous work indicated a single tryptic peptide difference between C4d.1 and C4d.2.

Purpose of the Study:

  • To define the specific amino acid difference underlying the C4d.1 and C4d.2 serologic specificities.
  • To investigate the molecular basis of H-2 associated alloantigenic variation in the complement system.

Main Methods:

  • Preparation and sequencing of genomic clones for C4d regions from H-2 haplotypes.
  • Comparison of C4d sequences from C4d.1 and C4d.2 types.
  • Analysis of restriction fragment length polymorphisms (RFLP) using Pst I and HindIII.

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Main Results:

  • The serologic difference between C4d.1 and C4d.2 is attributed to a single amino acid substitution: glutamine in C4d.1 versus arginine in C4d.2.
  • This substitution occurs in a hydrophilic region of the C4 molecule, homologous to a site involved in human C4 Chido/Rodgers differences.
  • A new Pst I site is identified in C4d.1 strains, and HindIII RFLP is observed between C4d.1 and C4d.2.

Conclusions:

  • A single amino acid substitution explains the distinct serological phenotypes of C4d.1 and C4d.2.
  • The identified substitution site is functionally significant, influencing both alloantigenicity and restriction enzyme recognition.
  • Molecular analysis provides a precise definition of alloantigenic variation within the complement C4 system.