SYNTHESIS, ASSEMBLY, AND SECRETION OF GAMMA GLOBULIN BY MOUSE MYELOMA CELLS

Abstract

MPC-11 myeloma tumor cells were adapted to growth in continuous culture. The cultured cells resembled the parent tumor in that they produced the fully assembled gamma globulin molecules as well as six unassembled molecules. Although cultured and tumor cells synthesized excess light chains, the molar ratio of light (L) to heavy (H) chains was approximately 1.7:1 in the culture, and 3.5:1 in the tumor. The cultured cells also produced fewer half molecules and free light chains than the parent tumor. Peptide column analysis did not reveal differences in the primary structure of the H chains derived from the parent tumor and the culture. The L chains may have differed by a minor peptide. As much as 20% of the newly labeled cytoplasmic proteins and almost 100% of the proteins secreted by the cultured myeloma cells could be precipitated by specific antiserum. The immune precipitates contained seven different gamma globulin molecules, six of which were characterized according to their molecular size and H and L chain content as fully assembled molecules (H2L2), heavy chain dimers (H2), half molecules (HL), H, light chain dimers (L2), and L chains. All gamma globulin subunits as well as the complete H2L2 molecule were produced and secreted by splenic clones of the parent MPC-11 tumor, and agar clones of the cultured cells. This indicates that the various gamma globulin subunits were produced by the same cell and did not reflect cellular heterogeneity with respect to gamma globulin synthesis.