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Sodium thiosulfate was used to enhance in vivo the polymerization of myeloma IgM, deficient in disulfide cross-links. The therapy sharply decreased the amount of low molecular weight IgM fractions, while increasing the serum content of molecules of higher molecular weight. The degree of disulfide cross-linking in IgM increased under the influence of thiosulfate. The rate of secretion into the serum and urine of some membrane-related glycopeptides and species rich in sialic acid was reduced. Also, the discharge of L chains to the urine was lowered during the thiosulfate trial. All these changes were attributed to enhancement of disulfide-interchanging enzyme activity by thiosulfate.

It has been suggested that the low disulfide interchange activity in cells producing IgM causes the excretion of immunoglobulin molecules with deficient disulfide cross-linking. IgM material found in the serum formed aggregates which were stabilized largely by noncovalent bonds. Sedimentation coefficients of these aggregates ranged from 11 to 19 S and were similar to sedimentation coefficients calculated for molecules composed of 2, 3, 4 and 5 IgM subunits. A fraction corresponding to a single IgM subunit with a sedimentation coefficient 7.1 S was also present in the serum. The molecular weight of this fraction, determined in mild conditions by Sephadex gel filtration, appeared higher than that predicted for IgMs, reaching the value of 270 000-280 000. The evident increment of molecular weight, compared to a single IgM subunit, was attributed to glycopeptides of sedimentation coefficient 2 S, which form the complex with IgMs molecules. These glycopeptides were also present in aggregates consisting of more than one IgM subunit. The number of IgM subunits participating in aggregates and their molecular weights appeared to depend on the ratio of 2 S glycopeptides to IgM material. With the increase of this ratio the molecular weight of the aggregates lowered and vice versa. Electrical charge and some physico-chemical properties of the aggregates were also significantly affected by the presence of glycopeptides, 2 S glycopeptides were supposed to be the basement membrane related material. Suggestions concerning the interpretation of the phenomenon are presented and discussed.

An acidic glycoprotein with calcium-binding properties was isolated from the urine of patients with severe macroglobulinaemia IgM. The molecular weight of this protein determined by Sephadex gel filtration was found to be 62 000 +/- 2800 in Tris - HCl buffer and 21 000 +/- 1000 in 6 M guanidine - HCl. The amino acid and carbohydrate composition of the isolated glycoprotein is presented. Electrophoretic migration of this protein was observed to be greatly affected by calcium ions present in the buffer in a concentration of 10(-3) M. At least two sets of binding sites seem to participate in binding calcium. The values 2.2 - 10(6) M-1 for the apparent association constant and 4.4 - 10(-4) mol of Ca2+ bound per g of protein for high affinity bindings sites were estimated, on the basis of data from the equilibrium dialysis. The origin and possible biological role of this protein is discussed.

Excessive production of IgM in macroglobulinemia is accompanied frequently by secretion of proteins and peptides which are normally absent from biological fluids. These proteins and peptides are probably secreted by cells producing IgM. Three types of peptides were isolated and their basic properties are described. One peptide showed calcium-binding properties, and the other two peptides were bound to secreted IgM immunoglobulin.