Phage display and peptide mapping of an immunoglobulin light chain fibril-related conformational epitope

Brian O'Nuallain, Amy Allen, Demet Ataman, Deborah T. Weiss, Alan Solomon, Jonathan Wall

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Amyloid fibrils and partially unfolded intermediates can be distinguished serologically from native amyloidogenic precursor proteins or peptides. In this regard, we previously had reported that mAb 11-1F4, generated by immunizing mice with a thermally denatured variable domain (VL) fragment of the human κ4 Bence Jones protein Len, bound to a non-native conformational epitope located within the N-terminal 18 residues of fibrillar, as well as partially denatured, Ig light chains (O'Nuallain, B., et al. (2006) Biochemistry 46, 1240-1247). To define further the antibody binding site, we used random peptide phage display and epitope mapping of VL Len using wild-type and alanine-mutated Len peptides where it was shown that the antibody epitope was reliant on up to 10 of the first 15 residues of protein Len. Comparison of Wκ and Vλ N-terminal germline consensus sequences with protein Len and 11-1F4-binding phages indicated that this antibody's cross-reactivity with light chains was related to an invariant proline at position(s) 7 and/or 8, bulky hydrophobic residues at positions 11 and 13, and additionally, to the ability to accommodate amino acid diversity at positions 1-4. Sequence alignments of the phage peptides revealed a central proline, often flanked by aromatic residues. Taken together, these results have provided evidence for the structural basis of the specificity of 11-1F4 for both κ and λ light chain fibrils. We posit that the associated binding site involves a rare type VI β-turn or touch-turn that is anchored by a cis-proline residue. The identification of an 11-1F4-related mimotope should facilitate development of pan-light chain fibril-reactive antibodies that could be used in the diagnosis and treatment of patients with AL amyloidosis.

Original languageEnglish (US)
Pages (from-to)13049-13058
Number of pages10
JournalBiochemistry
Volume46
Issue number45
DOIs
StatePublished - Nov 13 2007

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Immunoglobulin Light Chains
Bacteriophages
Peptide Mapping
Epitopes
Display devices
Proline
Light
Peptides
Antibodies
Bence Jones Protein
Antibody Binding Sites
Epitope Mapping
Amyloidogenic Proteins
Biochemistry
Aptitude
Protein Precursors
Sequence Alignment
Consensus Sequence
Touch
Amyloidosis

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Phage display and peptide mapping of an immunoglobulin light chain fibril-related conformational epitope. / O'Nuallain, Brian; Allen, Amy; Ataman, Demet; Weiss, Deborah T.; Solomon, Alan; Wall, Jonathan.

In: Biochemistry, Vol. 46, No. 45, 13.11.2007, p. 13049-13058.

Research output: Contribution to journalArticle

O'Nuallain, Brian ; Allen, Amy ; Ataman, Demet ; Weiss, Deborah T. ; Solomon, Alan ; Wall, Jonathan. / Phage display and peptide mapping of an immunoglobulin light chain fibril-related conformational epitope. In: Biochemistry. 2007 ; Vol. 46, No. 45. pp. 13049-13058.
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