Localization of a conformational epitope common to non-native and fibrillar immunoglobulin light chains

Brian O'Nuallain, Amy Allen, Stephen Kennel, Deborah T. Weiss, Alan Solomon, Jonathan Wall

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Amyloid fibrils and partially unfolded intermediates may be distinguished serologically from native amyloidogenic precursor proteins or peptides. In this regard, we had previously reported that the IgG1 mAb 11-1F4, generated by immunizing mice with a thermally denatured variable region fragment of the human Igκ4 Bence Jones protein Len, reacted specifically with light chain (LC) fibrils, irrespective of κ or λ isotype but, notably, did not with native molecules (Hrncic, R. et al. (2000) Am. J. Pathol. 157, 1239-1246). To elucidate the molecular basis of this specificity, we have used a europium-linked fluorescent immunoassay, where it was demonstrated through epitope mapping that mAb 11-1F4 recognizes a conformational determinant contained within the first (N-terminal) 18 amino acids of misfolded LCs. The nature of this epitope was evidenced in competition studies where the peptide Len (1-18), but not the intact protein or other LCs, inhibited the binding of the antibody to fibrils. This unique reactivity was dependent on the structural integrity of this portion of the molecule, particularly the presence of a highly conserved prolyl residue at position 8. On the basis of our experimental data, we posit that the mAb 11-1F4 binding site found on partially denatured and fibrillar LCs involves an inducible N-terminal main chain reversal that results in the formation of a proline anchored β-turn. Our delineation of this LC fibril-associated epitope provides the rationale for the design of novel amyloid-reactive antibodies with diagnostic and therapeutic potential for patients with LC-associated and other forms of amyloidosis.

Original languageEnglish (US)
Pages (from-to)1240-1247
Number of pages8
JournalBiochemistry
Volume46
Issue number5
DOIs
StatePublished - Feb 6 2007

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Immunoglobulin Light Chains
Epitopes
Light
Amyloid
Bence Jones Protein
Europium
Epitope Mapping
Amyloidogenic Proteins
Peptides
Molecules
Protein Precursors
Antibodies
Structural integrity
Amyloidosis
Immunoassay
Proline
Immunoglobulin G
Binding Sites
Amino Acids
Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Localization of a conformational epitope common to non-native and fibrillar immunoglobulin light chains. / O'Nuallain, Brian; Allen, Amy; Kennel, Stephen; Weiss, Deborah T.; Solomon, Alan; Wall, Jonathan.

In: Biochemistry, Vol. 46, No. 5, 06.02.2007, p. 1240-1247.

Research output: Contribution to journalArticle

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