Residues 484-508 Contain a Major Determinant of the Inhibitory Epitope in the A2 Domain of Human Factor VIII (*)

The A2 domain (residues 373-740) of human blood coagulation factor VIII (fVIII) contains a major epitope for inhibitory alloantibodies and autoantibodies. We took advantage of the differential reactivity of inhibitory antibodies with human and porcine fVIII and mapped a major determinant of the A2 epitope by using a series of active recombinant hybrid human/porcine fVIII molecules. Hybrids containing a substitution of porcine sequence at segment 410-508, 445-508, or 484-508 of the human A2 domain were not inhibited by a murine monoclonal antibody A2 inhibitor, mAb 413, whereas hybrids containing substitutions at 387-403, 387-444, and 387-468 were inhibited by mAb 413. This indicates that the segment bounded by Arg484 and Ile508 contains a major determinant of the A2 epitope. mAb 413 did not inhibit two more hybrids that contained porcine substitutions at residues 484-488 and 489-508, indicating that amino acid side chains on both sides of the Ser488-Arg489 bond within the Arg484-Ile508 segment contribute to the A2 epitope. The 484-508, 484-488, and 489-508 porcine substitution hybrids displayed decreased inhibition by A2 inhibitors from four patient plasmas, suggesting that there is little variation in the structure of the A2 epitope in the inhibitor population.

• Publication year

  1995

• Venue

  Journal of Biological Chemistry

• Publication date

  1995-06-16

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1074/JBC.270.24.14505PMID 7540171
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• Amino acid side chains on both sides of the Ser488-Arg489 bond contribute to the A2 epitope, and hybrids with 484-508, 484-488, or 489-508 porcine substitutions showed decreased inhibition by A2 inhibitors from four patient plasmas.

  Confidence 0.95

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewmexicorea (qjvnbu8xg3) reviewq (76h6bfydm6) review
• The segment bounded by Arg484 and Ile508 contains a major determinant of the A2 epitope.

  Confidence 0.97

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewmexicorea (qjvnbu8xg3) reviewq (76h6bfydm6) review
• Porcine substitutions spanning 410-508, 445-508, or 484-508 removed inhibition by mAb 413, whereas substitutions spanning 387-403, 387-444, or 387-468 did not.

  Confidence 0.98

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewmexicorea (qjvnbu8xg3) reviewq (76h6bfydm6) review

• a2 epitope

  epitope, antigenic site

  The inhibitory antibody-binding site within the A2 domain of human factor VIII examined in this work.

  Aliases: A2 inhibitory epitope

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewmexicorea (qjvnbu8xg3) reviewq (76h6bfydm6) review
• a2 inhibitors

  antibody, inhibitor

  Inhibitory antibodies in patient plasma that act against the A2 domain of factor VIII in the inhibition assays.

  Aliases: inhibitory antibodies, A2-domain inhibitors

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewmexicorea (qjvnbu8xg3) reviewq (76h6bfydm6) review
• arg484-ile508 segment

  protein segment, domain region

  The A2-domain segment of factor VIII defined by residues 484 through 508 and examined as a candidate epitope determinant.

  Aliases: residues 484-508 segment, 484-508 segment

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewmexicorea (qjvnbu8xg3) reviewq (76h6bfydm6) review
• mab 413

  antibody, experimental reagent

  A murine monoclonal antibody used as an A2-domain inhibitor probe in the hybrid factor VIII mapping experiments.

  Aliases: murine monoclonal antibody A2 inhibitor, monoclonal antibody 413

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewmexicorea (qjvnbu8xg3) reviewq (76h6bfydm6) review
• porcine sequence substitution hybrids

  protein construct, recombinant molecule

  Recombinant human/porcine factor VIII hybrid molecules carrying porcine sequence replacements in selected A2-domain segments.

  Aliases: human/porcine fVIII hybrids, hybrid human/porcine factor VIII molecules

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewmexicorea (qjvnbu8xg3) reviewq (76h6bfydm6) review
• ser488-arg489 bond

  chemical bond, protein feature

  The peptide bond between Ser488 and Arg489 within the A2-domain segment that was tested for epitope contribution.

  Aliases: Ser488-Arg489 peptide bond

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) reviewmexicorea (qjvnbu8xg3) reviewq (76h6bfydm6) review

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