Inhibition of the classical complement pathway by synthetic peptides from the second constant domain of the heavy chain of human immunoglobulin G

Michael B. Prystowsky, J. Michael Kehoe, Bruce W. Erickson

Research output: Contribution to journalArticle

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Abstract

The Cγ2 domain of immunoglobulin G (IgG) is reported to have a tryptophan residue and cationic residues at or near its C1q-binding site. The present study has used synthetic IgG peptides to explore the involvement of the 275-290 region of the Cγ2 domain of the heavy chain of human IgG1 in binding to C1. This region (Phe-Asn-Trp-Tyr-Val-Asp-Gly-Val-Gln-Val-His-Asn-Ala-Lys-Thr-Lys) contains Trp-277 and the cationic residues His-285, Lys-288, and Lys-290. The following peptides were synthesized by the solid-phase method and purified to homogeneity by using reverse-phase high-pressure liquid chromatography: the hexadecapeptide 275-290 and its Ni-formyl derivative 275-290F containing both Trp-277 and the cationic residues; the Nα-acetylpentapeptide 275-279A comprising the hydrophobic region around Trp-277; and the cationic decapeptide 281-290. When examined in the Augener assay for inhibition of C1-mediated immune hemolysis, peptides 275-290F and 281-290 were about half as active as monomeric 7S human IgG on a molar basis and essentially as active on a site basis. Since both peptides containing residues 281-290 inhibited hemolysis in a manner similar to the Cγ2 domain, the cationic 281-290 region containing His-285, Lys-288, and Lys-290 may be a part of the C1q-binding site of Cγ2. These results are con-sistent with the tertiary structure of the Fc fragment of IgG, in which the 275-279 region is part of the hydrophobic core of the Cγ2 domain and the 281-290 region is exposed on the surface.

Original languageEnglish (US)
Pages (from-to)6349-6356
Number of pages8
JournalBiochemistry
Volume20
Issue number22
StatePublished - 1981
Externally publishedYes

Fingerprint

Classical Complement Pathway
Immunoglobulin Heavy Chains
Immunoglobulin G
Peptides
Hemolysis
Binding Sites
High pressure liquid chromatography
Immunoglobulin Fc Fragments
Reverse-Phase Chromatography
Tryptophan
Assays
High Pressure Liquid Chromatography
Derivatives
C2 Domains

ASJC Scopus subject areas

  • Biochemistry

Cite this

Inhibition of the classical complement pathway by synthetic peptides from the second constant domain of the heavy chain of human immunoglobulin G. / Prystowsky, Michael B.; Kehoe, J. Michael; Erickson, Bruce W.

In: Biochemistry, Vol. 20, No. 22, 1981, p. 6349-6356.

Research output: Contribution to journalArticle

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N2 - The Cγ2 domain of immunoglobulin G (IgG) is reported to have a tryptophan residue and cationic residues at or near its C1q-binding site. The present study has used synthetic IgG peptides to explore the involvement of the 275-290 region of the Cγ2 domain of the heavy chain of human IgG1 in binding to C1. This region (Phe-Asn-Trp-Tyr-Val-Asp-Gly-Val-Gln-Val-His-Asn-Ala-Lys-Thr-Lys) contains Trp-277 and the cationic residues His-285, Lys-288, and Lys-290. The following peptides were synthesized by the solid-phase method and purified to homogeneity by using reverse-phase high-pressure liquid chromatography: the hexadecapeptide 275-290 and its Ni-formyl derivative 275-290F containing both Trp-277 and the cationic residues; the Nα-acetylpentapeptide 275-279A comprising the hydrophobic region around Trp-277; and the cationic decapeptide 281-290. When examined in the Augener assay for inhibition of C1-mediated immune hemolysis, peptides 275-290F and 281-290 were about half as active as monomeric 7S human IgG on a molar basis and essentially as active on a site basis. Since both peptides containing residues 281-290 inhibited hemolysis in a manner similar to the Cγ2 domain, the cationic 281-290 region containing His-285, Lys-288, and Lys-290 may be a part of the C1q-binding site of Cγ2. These results are con-sistent with the tertiary structure of the Fc fragment of IgG, in which the 275-279 region is part of the hydrophobic core of the Cγ2 domain and the 281-290 region is exposed on the surface.

AB - The Cγ2 domain of immunoglobulin G (IgG) is reported to have a tryptophan residue and cationic residues at or near its C1q-binding site. The present study has used synthetic IgG peptides to explore the involvement of the 275-290 region of the Cγ2 domain of the heavy chain of human IgG1 in binding to C1. This region (Phe-Asn-Trp-Tyr-Val-Asp-Gly-Val-Gln-Val-His-Asn-Ala-Lys-Thr-Lys) contains Trp-277 and the cationic residues His-285, Lys-288, and Lys-290. The following peptides were synthesized by the solid-phase method and purified to homogeneity by using reverse-phase high-pressure liquid chromatography: the hexadecapeptide 275-290 and its Ni-formyl derivative 275-290F containing both Trp-277 and the cationic residues; the Nα-acetylpentapeptide 275-279A comprising the hydrophobic region around Trp-277; and the cationic decapeptide 281-290. When examined in the Augener assay for inhibition of C1-mediated immune hemolysis, peptides 275-290F and 281-290 were about half as active as monomeric 7S human IgG on a molar basis and essentially as active on a site basis. Since both peptides containing residues 281-290 inhibited hemolysis in a manner similar to the Cγ2 domain, the cationic 281-290 region containing His-285, Lys-288, and Lys-290 may be a part of the C1q-binding site of Cγ2. These results are con-sistent with the tertiary structure of the Fc fragment of IgG, in which the 275-279 region is part of the hydrophobic core of the Cγ2 domain and the 281-290 region is exposed on the surface.

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