Amino acid residues 1101-1105 of the isotypic region of human C4B is important to the covalent binding activity of complement component C4.
The C4A and C4B isotypes of human C4 show certain functional differences that stem from their relative preference for transacylation to amino (-NH2) vs hydroxyl (-OH) nucleophiles, respectively, on complement-activating surfaces. Comparison of amino acid sequences of the alpha-chain fragment of C4, C4d, has shown C4A- and C4B-specific sequences at residues 1101-1106 are the only consistent structural difference between isotype, i.e., Pro, Cys, Pro, Val, Leu, Asp in C4A and Leu, Ser, Pro, Val Ile, His in C4B. These residues may be responsible either in part or entirely for properties associated with isotype. To examine the functional role of residues 1101-1106 in C4B-mediated hemolysis, whole serum or immunopurified human C4 with allotypes, A3B1, A3, B2B1, or B1 were preincubated in the presence or absence of an antipeptide mAb (BII-1) specific for amino acid residues 1101-1105 of C4B. Sensitized sheep E and C4-deficient guinea pig serum was then added and lysis measured by absorbance at 415 nm. Our results show lysis of antibody-sensitized sheep E is inhibited by antibody and C4B2B1, C4B1, or C4A3B1 but not antibody and C4A3. The interference of hemolysis by BII-1 could not be explained by inhibition of activation of C4B or inhibition of C3 or C5 convertase activity. Furthermore, results from uptake experiments show that BII-1 interferes with the covalent binding activity of C4B, indicating residues 1101-1105 play a role in the covalent binding reaction of C4B to the target E-antibody complex.
Faculty of Medicine, Memorial University of Newfoundland, St. John's, Canada.
B D
BD
Reilly
R P
RP
Levine
V M
VM
Skanes
D000595:Amino Acid Sequence; D000906:Antibodies / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000276:immunology; D003167:Complement Activation; D003173:Complement C1s / Q000378:metabolism; D050577:Complement C3-C5 Convertases / Q000378:metabolism; D015935:Complement C4b / Q000737:chemistry / Q000276:immunology*; D006461:Hemolysis; D006801:Humans; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000276:immunology
D000906:Antibodies; D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D010455:Peptides; D015935:Complement C4b; D050577:Complement C3-C5 Convertases; D003173:Complement C1s