Assembly, secretion, and vacuolar delivery of a hybrid immunoglobulin in plants

Plant Physiol. 2000 Aug;123(4):1483-94. doi: 10.1104/pp.123.4.1483.

Abstract

Secretory immunoglobulin (Ig) A is a decameric Ig composed of four alpha-heavy chains, four light chains, a joining (J) chain, and a secretory component (SC). The heavy and light chains form two tetrameric Ig molecules that are joined by the J chain and associate with the SC. Expression of a secretory monoclonal antibody in tobacco (Nicotiana tabacum) has been described: this molecule (secretory IgA/G [SIgA/G]) was modified by having a hybrid heavy chain sequence consisting of IgG gamma-chain domains linked to constant region domains of an IgA alpha-chain. In tobacco, about 70% of the protein assembles to its final, decameric structure. We show here that SIgA/G assembly and secretion are slow, with only approximately 10% of the newly synthesized molecules being secreted after 24 h and the bulk probably remaining in the endoplasmic reticulum. In addition, a proportion of SIgA/G is delivered to the vacuole as at least partially assembled molecules by a process that is blocked by the membrane traffic inhibitor brefeldin A. Neither the SC nor the J chain are responsible for vacuolar delivery, because IgA/G tetramers have the same fate. The parent IgG tetrameric molecule, containing wild-type gamma-heavy chains, is instead secreted rapidly and efficiently. This strongly suggests that intracellular retention and vacuolar delivery of IgA/G is due to the alpha-domains present in the hybrid alpha/gamma-heavy chains and indicates that the plant secretory system may partially deliver to the vacuole recombinant proteins expected to be secreted.

MeSH terms

  • Brefeldin A / pharmacology
  • Immunoglobulin A, Secretory / genetics*
  • Immunoglobulin A, Secretory / metabolism
  • Immunoglobulin G / genetics*
  • Immunoglobulin G / metabolism
  • Immunohistochemistry
  • Microscopy, Confocal
  • Microscopy, Immunoelectron
  • Nicotiana / genetics*
  • Nicotiana / metabolism
  • Nicotiana / ultrastructure
  • Plant Leaves / metabolism
  • Plants, Toxic*
  • Precipitin Tests
  • Protein Synthesis Inhibitors / pharmacology
  • Recombinant Fusion Proteins / genetics*
  • Recombinant Fusion Proteins / metabolism
  • Vacuoles / metabolism*
  • Vacuoles / ultrastructure

Substances

  • Immunoglobulin A, Secretory
  • Immunoglobulin G
  • Protein Synthesis Inhibitors
  • Recombinant Fusion Proteins
  • Brefeldin A