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. 1968 Jul;108(3):375–382. doi: 10.1042/bj1080375

Specific dimerization of the light chains of human immunoglobulin

G T Stevenson 1, D Straus 1,*
PMCID: PMC1198822  PMID: 4174431

Abstract

1. The light chains of human immunoglobulin were allowed to dimerize in vitro on removal of the dispersing agents acetic acid or urea. 2. On electrophoresis in polyacrylamide gel at pH8·8 the dimers yielded up to nine regularly spaced bands. This approximates to the number of electrophoretic components known to occur among the monomers. 3. Single electrophoretic components of the dimers were isolated from the gel, dissociated into monomers, and subjected as such to electrophoresis in urea-containing gels. Each gave two adjacent bands. 4. Similarly, after all the light chains as monomers had been subjected to electrophoresis in urea-containing gels, single electrophoretic components were isolated and allowed to dimerize. When examined now as dimers in the absence of urea, each component gave two adjacent bands. 5. These findings are explicable on the following basis. (a) The dimerization of the light chains is specific, at least inasmuch as it occurs between monomers of the same electrophoretic mobilities. (b) With the buffer constant, different light chains undergo different changes in net charge on being transferred from urea-containing to urea-free solution; in this way two different chains of the same initial charge can acquire a charge difference of 1. 6. Experiments with Bence-Jones proteins and other homogeneous light chains gave results substantiating the conclusions (a) and (b).

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Brewer J. M. Artifact produced in disc electrophoresis by ammonium persulfate. Science. 1967 Apr 14;156(3772):256–257. doi: 10.1126/science.156.3772.256. [DOI] [PubMed] [Google Scholar]
  2. Cohen S., Gordon S. Dissociation of kappa- and lambda-chains from reduced human immunoglobulins. Biochem J. 1965 Nov;97(2):460–465. doi: 10.1042/bj0970460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cohen S., Milstein C. Structure and biological properties of immunoglobulins. Adv Immunol. 1967;7:1–89. doi: 10.1016/s0065-2776(08)60126-1. [DOI] [PubMed] [Google Scholar]
  4. Cohen S., Porter R. R. Heterogeneity of the peptide chains of gamma-globulin. Biochem J. 1964 Feb;90(2):278–284. doi: 10.1042/bj0900278. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  6. FERRIS T. G., EASTERLING R. E., BUDD R. E. Electrophoresis of serum proteins in acrylamide gel. I. Removal of background stain by means of electric current. Am J Clin Pathol. 1962 Oct;38:383–387. doi: 10.1093/ajcp/38.4.383. [DOI] [PubMed] [Google Scholar]
  7. FLEISCHMAN J. B., PAIN R. H., PORTER R. R. Reduction of gamma-globulins. Arch Biochem Biophys. 1962 Sep;Suppl 1:174–180. [PubMed] [Google Scholar]
  8. FLEISCHMAN J. B., PORTER R. R., PRESS E. M. THE ARRANGEMENT OF THE PEPTIDE CHAINS IN GAMMA-GLOBULIN. Biochem J. 1963 Aug;88:220–228. doi: 10.1042/bj0880220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Feinstein A. Use of charged thiol reagents in interpreting the electrophoretic patterns of immune globulin chains and fragments. Nature. 1966 Apr 9;210(5032):135–137. doi: 10.1038/210135a0. [DOI] [PubMed] [Google Scholar]
  10. GALLY J. A., EDELMAN G. M. PROTEIN-PROTEIN INTERACTIONS AMONG L POLYPEPTIDE CHAINS OF BENCE-JONES PROTEINS AND HUMAN GAMMA-GLOBULINS. J Exp Med. 1964 May 1;119:817–836. doi: 10.1084/jem.119.5.817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hilschmann N., Craig L. C. Amino acid sequence studies with Bence-Jones proteins. Proc Natl Acad Sci U S A. 1965 Jun;53(6):1403–1409. doi: 10.1073/pnas.53.6.1403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Martin R. G., Goldberger R. F. Imidazolylacetolphosphate:L-glutamate aminotransferase. Purification and physical properties. J Biol Chem. 1967 Mar 25;242(6):1168–1174. [PubMed] [Google Scholar]
  13. Milstein C. Variations in amino-acid sequence near the disulphide bridges of Bence-Jones proteins. Nature. 1966 Jan 22;209(5021):370–373. doi: 10.1038/209370a0. [DOI] [PubMed] [Google Scholar]
  14. Nachman R. L., Engle R. L., Jr, Stein S. Subtypes of Bence-Jones proteins. J Immunol. 1965 Aug;95(2):295–299. [PubMed] [Google Scholar]
  15. ORNSTEIN L. DISC ELECTROPHORESIS. I. BACKGROUND AND THEORY. Ann N Y Acad Sci. 1964 Dec 28;121:321–349. doi: 10.1111/j.1749-6632.1964.tb14207.x. [DOI] [PubMed] [Google Scholar]
  16. PAIN R. H. THE MOLECULAR WEIGHTS OF THE PEPTIDE CHAINS OF GAMMA-GLOBULIN. Biochem J. 1963 Aug;88:234–239. doi: 10.1042/bj0880234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Poulik M. D. Gel electrophoresis in buffers containing urea. Methods Biochem Anal. 1966;14:455–495. doi: 10.1002/9780470110324.ch9. [DOI] [PubMed] [Google Scholar]
  18. RAYMOND S. A convenient apparatus for vertical gel electrophoresis. Clin Chem. 1962 Sep-Oct;8:455–470. [PubMed] [Google Scholar]
  19. SMALL P. A., Jr, REISFELD R. A., DRAY S. PEPTIDE DIFFERENCES OF RABBIT GAMMA-G-IMMUNOGLOBULIN LIGHT CHAINS CONTROLLED BY ALLELIC GENES. J Mol Biol. 1965 Apr;11:713–721. doi: 10.1016/s0022-2836(65)80029-8. [DOI] [PubMed] [Google Scholar]
  20. SMITHIES O. Zone electrophoresis in starch gels and its application to studies of serum proteins. Adv Protein Chem. 1959;14:65–113. doi: 10.1016/s0065-3233(08)60609-9. [DOI] [PubMed] [Google Scholar]
  21. STEVENSON G. T. Further studies of the gamma-related proteins of normal urine. J Clin Invest. 1962 May;41:1190–1198. doi: 10.1172/JCI104572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Shapiro A. L., Scharff M. D., Maizel J. V., Jr, Uhr J. W. Polyribosomal synthesis and assembly of the H and L chains of gamma globulin. Proc Natl Acad Sci U S A. 1966 Jul;56(1):216–221. doi: 10.1073/pnas.56.1.216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shapiro A. L., Scharff M. D., Maizel J. V., Uhr J. W. Synthesis of excess light chains of gamma globulin by rabbit lymph node cells. Nature. 1966 Jul 16;211(5046):243–245. doi: 10.1038/211243a0. [DOI] [PubMed] [Google Scholar]
  24. Titani K., Whitley E., Jr, Putnam F. W. Immunoglobulin structure: variation in the sequence of Bence Jones proteins. Science. 1966 Jun 10;152(3728):1513–1516. doi: 10.1126/science.152.3728.1513. [DOI] [PubMed] [Google Scholar]

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