Expression of the recombinant human immunoglobulin J chain in Escherichia coli

Mol Immunol. 2000 Feb-Mar;37(3-4):133-40. doi: 10.1016/s0161-5890(00)00035-3.

Abstract

Selective transport of polymeric (p) immunoglobulins (Ig) of IgA and IgM isotypes into external secretions by pIg receptor-mediated mechanism depends on the incorporation of joining (J) chain into the polymers. Until now, availability of a free J chain for immunological and biophysical studies has been limited to preparations of denatured J chain forms with moderate yield. Here we report that a recombinant J chain (rJ) can be over-expressed as a soluble fusion protein with thioredoxin using a modified vector pET32 in Escherichia coli. An intact J chain was released by digestion with IgA1 protease from Neisseria gonorrhoeae and isolated in a good yield with immunological and biochemical properties similar to those of J chain obtained by chemical cleavage from pIgA.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Humans
  • Immunoglobulin A
  • Immunoglobulin J-Chains / biosynthesis*
  • Immunoglobulin J-Chains / genetics
  • Immunoglobulin M
  • Molecular Sequence Data
  • Recombinant Fusion Proteins / biosynthesis*
  • Serine Endopeptidases / metabolism
  • Thioredoxins / biosynthesis
  • Thioredoxins / genetics

Substances

  • Immunoglobulin A
  • Immunoglobulin J-Chains
  • Immunoglobulin M
  • Recombinant Fusion Proteins
  • polymeric IgA
  • polymeric IgM
  • Thioredoxins
  • Serine Endopeptidases
  • IgA-specific serine endopeptidase