Mutation of a single conserved residue in VH complementarity-determining region 2 results in a severe Ig secretion defect

G. D. Wiens, A. Lekkerkerker, I. Veltman, Marvin Rittenberg

Research output: Contribution to journalArticle

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Abstract

During an immune response, somatic mutations are introduced into the VH and VL regions of Ig chains. The consequences of somatic mutation in highly conserved residues are poorly understood. Ile51 is present in 91% of murine VH complementarity-determining region 2 sequences, and we demonstrate that single Ile51→Arg or Lys substitutions in the PCG1-1 Ab are sufficient to severely reduce Ig secretion (1-3% of wild-type (WT) levels). Mutant H chains, expressed in the presence of excess L chain, associate with Ig binding protein (BiP) and GRP94 and fail to form HL and H2L assembly intermediates efficiently. The mutations do not irreversibly alter the VH domain as the small amount of mutant H chain, which assembles with L chain as H2L2, is secreted. The secreted mutant Ab binds phosphocholine-protein with avidity identical with that of WT Ab, suggesting that the combining site adopts a WT conformation. A computer-generated model of the PCG1-1 variable region fragment of Ig (Fv) indicates that Ile51 is buried between complementarity-determining region 2 and framework 3 and does not directly contact the L chain. Thus, the Ile51→Arg or Ile51→Lys mutations impair association with the PCG1-1 L chain via indirect interactions. These interactions are in part dependent on the nature of the L chain as the PCG1-1 VH single Ile51→Arg or Ile51→Lys mutants were partially rescued when expressed with the J558L λ1 L chain. These results represent the first demonstration that single somatic mutations in Va residues can impair Ig secretion and suggest one reason for the conservation of Ile51 in so many Ig VH.

Original languageEnglish (US)
Pages (from-to)2179-2186
Number of pages8
JournalJournal of Immunology
Volume167
Issue number4
StatePublished - Aug 15 2001

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Complementarity Determining Regions
Mutation
Phosphorylcholine
Computer Simulation
Carrier Proteins
Binding Sites
Proteins

ASJC Scopus subject areas

  • Immunology

Cite this

Mutation of a single conserved residue in VH complementarity-determining region 2 results in a severe Ig secretion defect. / Wiens, G. D.; Lekkerkerker, A.; Veltman, I.; Rittenberg, Marvin.

In: Journal of Immunology, Vol. 167, No. 4, 15.08.2001, p. 2179-2186.

Research output: Contribution to journalArticle

Wiens, GD, Lekkerkerker, A, Veltman, I & Rittenberg, M 2001, 'Mutation of a single conserved residue in VH complementarity-determining region 2 results in a severe Ig secretion defect', Journal of Immunology, vol. 167, no. 4, pp. 2179-2186.
Wiens, G. D. ; Lekkerkerker, A. ; Veltman, I. ; Rittenberg, Marvin. / Mutation of a single conserved residue in VH complementarity-determining region 2 results in a severe Ig secretion defect. In: Journal of Immunology. 2001 ; Vol. 167, No. 4. pp. 2179-2186.
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