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. 1972 Jun;128(2):427–444. doi: 10.1042/bj1280427

The complete amino acid sequence of a mouse κ light chain

J Svasti 1, C Milstein 1
PMCID: PMC1173779  PMID: 4638343

Abstract

The complete amino acid sequence of the κ-chain of the mouse myeloma protein MOPC 21 was established. The protein was reduced and alkylated with iodo[2-14C]acetic acid, and 21 tryptic peptides were isolated, mainly by paper electrophoresis and paper chromatography. Three large tryptic peptides (of 35, 36 and 42 residues), which were difficult to isolate in this manner, were obtained pure and in excellent yields by a combination of Sephadex G-50 gel filtration in 1% (w/v) NH4HCO3 and chromatography on a DEAE-cellulose column in ammonium acetate buffer, pH8.1. Peptides overlapping the tryptic peptides were isolated from a chymotryptic digest. The chain is 214 residues long. Microheterogeneity of two peptides was observed and is believed to be due to deamidation. It was not excluded that such deamidation could occur in serum from which the protein was isolated. The sequence is compared with the sequences of two other mouse κ-chains, and with the human κ-chain basic sequences.

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Selected References

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

  1. AMBLER R. P. THE PURIFICATION AND AMINO ACID COMPOSITION OF PSEUDOMONAS CYTOCHROME C-551. Biochem J. 1963 Nov;89:341–349. doi: 10.1042/bj0890341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. ANFINSEN C. B., AQVIST S. E., COOKE J. P., JONSSON B. A comparative study of the structures of bovine and ovine pancreatic ribonucleases. J Biol Chem. 1959 May;234(5):1118–1123. [PubMed] [Google Scholar]
  3. Ambler R. P., Brown L. H. The amino acid sequence of Pseudomonas fluorescens azurin. Biochem J. 1967 Sep;104(3):784–825. doi: 10.1042/bj1040784. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Appella E. Amino acid sequences of two mouse immunoglobulin lambda chains. Proc Natl Acad Sci U S A. 1971 Mar;68(3):590–594. doi: 10.1073/pnas.68.3.590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Awdeh Z. L., Williamson A. R., Askonas B. A. One cell-one immunoglobulin. Origin of limited heterogeneity of myeloma proteins. Biochem J. 1970 Jan;116(2):241–248. doi: 10.1042/bj1160241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. BENNETT J. C., HOOD L. E., DREYER W. J., POTTER M. EVIDENCE FOR AMINO ACID SEQUENCE DIFFERENCES AMONG PROTEINS RESEMBLING THE L-CHAIN SUBUNITS OF IMMUNOGLOBULINS. J Mol Biol. 1965 May;12:81–87. doi: 10.1016/s0022-2836(65)80284-4. [DOI] [PubMed] [Google Scholar]
  7. Brown J. R., Hartley B. S. Location of disulphide bridges by diagonal paper electrophoresis. The disulphide bridges of bovine chymotrypsinogen A. Biochem J. 1966 Oct;101(1):214–228. doi: 10.1042/bj1010214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Crowshaw K., Jessup S. J., Ramwell P. W. Thin-layer chromatography of 1-dimethylaminonaphthalene-5-sulphonyl derivatives of amino acids present in superfusates of cat cerebral cortex. Biochem J. 1967 Apr;103(1):79–85. doi: 10.1042/bj1030079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Edman P., Begg G. A protein sequenator. Eur J Biochem. 1967 Mar;1(1):80–91. doi: 10.1007/978-3-662-25813-2_14. [DOI] [PubMed] [Google Scholar]
  11. Ein D. Nonallelic behavior of the Oz groups in human lambda immunoglobulin chains. Proc Natl Acad Sci U S A. 1968 Jul;60(3):982–985. doi: 10.1073/pnas.60.3.982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Frangione B., Milstein C., Franklin E. C. Chemical typing of immunoglobulins. Nature. 1969 Jan 11;221(5176):149–151. doi: 10.1038/221149a0. [DOI] [PubMed] [Google Scholar]
  13. Gibson D., Levanon M., Smithies O. Heterogeneity of normal human immunoglobulin light chains. Nonallelic variation in the constant region of lambda chains. Biochemistry. 1971 Aug 3;10(16):3114–3122. doi: 10.1021/bi00792a021. [DOI] [PubMed] [Google Scholar]
  14. Gray W. R., Dreyer W. J., Hood L. Mechanism of antibody synthesis: size differences between mouse kappa chains. Science. 1967 Jan 27;155(3761):465–467. doi: 10.1126/science.155.3761.465. [DOI] [PubMed] [Google Scholar]
  15. HEILMANN J., BARROLLIER J., WATZKE E. Beitrag zur Aminosäurebestimmung auf Papierchromatogrammen. Hoppe Seylers Z Physiol Chem. 1957;309(4-6):219–220. [PubMed] [Google Scholar]
  16. Hood L. E., Potter M., McKean D. J. Immunoglobulin structure: amino terminal sequences of kappa chains from genetically similar mice (BALB/c). Science. 1970 Dec 11;170(3963):1207–1210. doi: 10.1126/science.170.3963.1207. [DOI] [PubMed] [Google Scholar]
  17. Milstein C. P., Deverson E. V. The amino acid sequence of a human kappa light chain. Biochem J. 1971 Aug;123(5):945–958. doi: 10.1042/bj1230945. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Milstein C. A simple procedure for the fractionation of the tryptic peptides of the C-terminal half of immunoglobulin lambda-chains. Biochem J. 1968 Dec;110(4):652–654. doi: 10.1042/bj1100652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Milstein C. Linked groups of residues in immunoglobulin k chains. Nature. 1967 Oct 28;216(5113):330–332. doi: 10.1038/216330a0. [DOI] [PubMed] [Google Scholar]
  20. Milstein C., Munro A. J. The genetic basis of antibody specificity. Annu Rev Microbiol. 1970;24:335–358. doi: 10.1146/annurev.mi.24.100170.002003. [DOI] [PubMed] [Google Scholar]
  21. Milstein C., Pink J. R. Structure and evolution of immunoglobulins. Prog Biophys Mol Biol. 1970;21:209–263. doi: 10.1016/0079-6107(70)90026-x. [DOI] [PubMed] [Google Scholar]
  22. Milstein C. The disulphide bridges of immunoglobulin kappa-chains. Biochem J. 1966 Nov;101(2):338–351. doi: 10.1042/bj1010338. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Offord R. E. Electrophoretic mobilities of peptides on paper and their use in the determination of amide groups. Nature. 1966 Aug 6;211(5049):591–593. doi: 10.1038/211591a0. [DOI] [PubMed] [Google Scholar]
  24. Pink J. R., Buttery S. H., De Vries G. M., Milstein C. Human immunoglobulin subclasses. Partial amino acid sequence of the constant region of a gamma 4 chain. Biochem J. 1970 Mar;117(1):33–47. doi: 10.1042/bj1170033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Plummer T. H., Jr Isolation and sequence of peptides at the active center of bovine carboxypeptidase B. J Biol Chem. 1969 Oct 10;244(19):5246–5253. [PubMed] [Google Scholar]
  26. SANGER F., THOMPSON E. O. Halogenation of tyrosine during acid hydrolysis. Biochim Biophys Acta. 1963 May 14;71:468–471. doi: 10.1016/0006-3002(63)91108-9. [DOI] [PubMed] [Google Scholar]
  27. SMITH I. Colour reactions on paper chromatograms by a dipping technique. Nature. 1953 Jan 3;171(4340):43–44. doi: 10.1038/171043a0. [DOI] [PubMed] [Google Scholar]
  28. Schiechl H., Hilschmann N. Die Primärstruktur einer monoklonalen Immunglobulin-L-Kette der Subgruppe I vom kappa-Typ (Bence-Jones-Protein Au): gekoppelte Austausche innerhalb der Subgruppen. Hoppe Seylers Z Physiol Chem. 1971 Jan;352(1):111–115. [PubMed] [Google Scholar]
  29. Smith G. P., Hood L., Fitch W. M. Antibody diversity. Annu Rev Biochem. 1971;40:969–1012. doi: 10.1146/annurev.bi.40.070171.004541. [DOI] [PubMed] [Google Scholar]
  30. Svasti J., Milstein C. The disulphide bridges of a mouse immunoglobulin G1 protein. Biochem J. 1972 Feb;126(4):837–850. doi: 10.1042/bj1260837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. WALEY S. G., WATSON J. The action of trypsin on polylysine. Biochem J. 1953 Sep;55(2):328–337. doi: 10.1042/bj0550328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wang A. C., Pink J. R., Fudenberg H. H., Ohms J. A variable region subclass of heavy chains common to immunoglobulins G, A, and M and characterized by an unblocked amino-terminal residue. Proc Natl Acad Sci U S A. 1970 Jul;66(3):657–663. doi: 10.1073/pnas.66.3.657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Weigert M. G., Cesari I. M., Yonkovich S. J., Cohn M. Variability in the lambda light chain sequences of mouse antibody. Nature. 1970 Dec 12;228(5276):1045–1047. doi: 10.1038/2281045a0. [DOI] [PubMed] [Google Scholar]
  34. Woods K. R., Wang K. T. Separation of dansyl-amino acids by polyamide layer chromatography. Biochim Biophys Acta. 1967 Feb 21;133(2):369–370. doi: 10.1016/0005-2795(67)90078-5. [DOI] [PubMed] [Google Scholar]

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