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. 1985 Apr;82(8):2282–2286. doi: 10.1073/pnas.82.8.2282

Nucleotide sequence of cDNA encoding human alpha 2-macroglobulin and assignment of the chromosomal locus.

C C Kan, E Solomon, K T Belt, A C Chain, L R Hiorns, G Fey
PMCID: PMC397541  PMID: 2581245

Abstract

Six alpha 2-macroglobulin (alpha 2M) cDNA clones were isolated from a human liver cDNA library by using synthetic oligonucleotides as hybridization probes. One of these, p alpha 2M1, carries a 4.6 kilobase-pair insert, which was sequenced. The insert contains the coding sequences for the mature alpha 2M polypeptide (1451 amino acids) and for a 23-amino acid signal peptide at the NH2 terminus of the precursor pro-alpha 2M. At the 3' end of the insert a poly(A) addition signal A-A-T-A-A-A and part of the poly(A) tail of the messenger RNA were found. The protein sequence deduced from the nucleotide sequence agrees with the published alpha 2M amino acid sequence for all except three residues. The alpha 2M locus was assigned to human chromosome 12 by Southern blot analysis with DNA from a panel of mouse/human somatic cell hybrids, using alpha 2M cDNA as a hybridization probe.

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Selected References

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  1. Barrett A. J. Alpha 2-macroglobulin. Methods Enzymol. 1981;80(Pt 100):737–754. doi: 10.1016/s0076-6879(81)80056-0. [DOI] [PubMed] [Google Scholar]
  2. Barrett A. J., Starkey P. M. The interaction of alpha 2-macroglobulin with proteinases. Characteristics and specificity of the reaction, and a hypothesis concerning its molecular mechanism. Biochem J. 1973 Aug;133(4):709–724. doi: 10.1042/bj1330709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Belt K. T., Carroll M. C., Porter R. R. The structural basis of the multiple forms of human complement component C4. Cell. 1984 Apr;36(4):907–914. doi: 10.1016/0092-8674(84)90040-0. [DOI] [PubMed] [Google Scholar]
  4. Bernards R., Flavell R. A. Physical mapping of the globin gene deletion in hereditary persistence of foetal haemoglobin (HPFH). Nucleic Acids Res. 1980 Apr 11;8(7):1521–1534. doi: 10.1093/nar/8.7.1521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bobrow M., Cross J. Differential staining of human and mouse chromosomes in interspecific cell hybrids. Nature. 1974 Sep 6;251(5470):77–79. doi: 10.1038/251077a0. [DOI] [PubMed] [Google Scholar]
  7. Carroll M. C., Campbell R. D., Bentley D. R., Porter R. R. A molecular map of the human major histocompatibility complex class III region linking complement genes C4, C2 and factor B. Nature. 1984 Jan 19;307(5948):237–241. doi: 10.1038/307237a0. [DOI] [PubMed] [Google Scholar]
  8. Domdey H., Wiebauer K., Kazmaier M., Müller V., Odink K., Fey G. Characterization of the mRNA and cloned cDNA specifying the third component of mouse complement. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7619–7623. doi: 10.1073/pnas.79.24.7619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Doolittle R. F., Hunkapiller M. W., Hood L. E., Devare S. G., Robbins K. C., Aaronson S. A., Antoniades H. N. Simian sarcoma virus onc gene, v-sis, is derived from the gene (or genes) encoding a platelet-derived growth factor. Science. 1983 Jul 15;221(4607):275–277. doi: 10.1126/science.6304883. [DOI] [PubMed] [Google Scholar]
  10. Fey G. H., Lundwall A., Wetsel R. A., Tack B. F., de Bruijn M. H., Domdey H. Nucleotide sequence of complementary DNA and derived amino acid sequence of murine complement protein C3. Philos Trans R Soc Lond B Biol Sci. 1984 Sep 6;306(1129):333–344. doi: 10.1098/rstb.1984.0094. [DOI] [PubMed] [Google Scholar]
  11. Fey G. H., Wiebauer K., Domdey H. Amino acid sequences of mouse complement C3 derived from nucleotide sequences of cloned cDNA. Ann N Y Acad Sci. 1983;421:307–312. doi: 10.1111/j.1749-6632.1983.tb18118.x. [DOI] [PubMed] [Google Scholar]
  12. Fey G., Domdey H., Wiebauer K., Whitehead A. S., Odink K. Structure and expression of the C3 gene. Springer Semin Immunopathol. 1983;6(2-3):119–147. doi: 10.1007/BF00205869. [DOI] [PubMed] [Google Scholar]
  13. Goodfellow P. N., Banting G., Trowsdale J., Chambers S., Solomon E. Introduction of a human X-6 translocation chromosome into a mouse teratocarcinoma: investigation of control of HLA-A, B, C expression. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1190–1194. doi: 10.1073/pnas.79.4.1190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Goodfellow P. N., Banting G., Wiles M. V., Tunnacliffe A., Parkar M., Solomon E., Dalchau R., Fabre J. W. The gene, MIC4, which controls expression of the antigen defined by monoclonal antibody F10.44.2, is on human chromosome 11. Eur J Immunol. 1982 Aug;12(8):659–663. doi: 10.1002/eji.1830120807. [DOI] [PubMed] [Google Scholar]
  15. Gordon A. H. The alpha macroglobulins of rat serum. Biochem J. 1976 Dec 1;159(3):643–650. doi: 10.1042/bj1590643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hanover J. A., Willingham M. C., Pastan I. Receptor-mediated endocytosis of alpha 2-macroglobulin: solubilization and partial purification of the fibroblast alpha 2-macroglobulin receptor. Ann N Y Acad Sci. 1983;421:410–423. doi: 10.1111/j.1749-6632.1983.tb18135.x. [DOI] [PubMed] [Google Scholar]
  18. Harpel P. C., Brower M. S. Alpha 2-macroglobulin: an introduction. Ann N Y Acad Sci. 1983;421:1–9. doi: 10.1111/j.1749-6632.1983.tb18088.x. [DOI] [PubMed] [Google Scholar]
  19. Harpel P. C. Studies on human plasma alpha 2-macroglobulin-enzyme interactions. Evidence for proteolytic modification of the subunit chain structure. J Exp Med. 1973 Sep 1;138(3):508–521. doi: 10.1084/jem.138.3.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Howard J. B. Reactive centers in alpha 2-macroglobulin. Ann N Y Acad Sci. 1983;421:69–80. doi: 10.1111/j.1749-6632.1983.tb18093.x. [DOI] [PubMed] [Google Scholar]
  21. Huang J. S., Huang S. S., Deuel T. F. Specific covalent binding of platelet-derived growth factor to human plasma alpha 2-macroglobulin. Proc Natl Acad Sci U S A. 1984 Jan;81(2):342–346. doi: 10.1073/pnas.81.2.342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kaplan J., Keogh E. A. Studies on the physiology of macrophage receptors for alpha-macroglobulin X protease complexes. Ann N Y Acad Sci. 1983;421:442–456. doi: 10.1111/j.1749-6632.1983.tb18138.x. [DOI] [PubMed] [Google Scholar]
  23. Kushner I. The phenomenon of the acute phase response. Ann N Y Acad Sci. 1982;389:39–48. doi: 10.1111/j.1749-6632.1982.tb22124.x. [DOI] [PubMed] [Google Scholar]
  24. Minowada J., Onuma T., Moore G. E. Rosette-forming human lymphoid cell lines. I. Establishment and evidence for origin of thymus-derived lymphocytes. J Natl Cancer Inst. 1972 Sep;49(3):891–895. [PubMed] [Google Scholar]
  25. Nabholz M., Miggiano V., Bodmer W. Genetic analysis with human--mouse somatic cell hybrids. Nature. 1969 Jul 26;223(5204):358–363. doi: 10.1038/223358a0. [DOI] [PubMed] [Google Scholar]
  26. Nonaka M., Takahashi M., Natsuume-Sakai S., Nonaka M., Tanaka S., Shimizu A., Honjo T. Isolation of cDNA clones specifying the fourth component of mouse complement and its isotype, sex-limited protein. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6822–6826. doi: 10.1073/pnas.81.21.6822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ogata R. T., Shreffler D. C., Sepich D. S., Lilly S. P. cDNA clone spanning the alpha-gamma subunit junction in the precursor of the murine fourth complement component (C4). Proc Natl Acad Sci U S A. 1983 Aug;80(16):5061–5065. doi: 10.1073/pnas.80.16.5061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sim R. B., Sim E. Autolytic fragmentation of complement components C3 and C4 under denaturing conditions, a property shared with alpha 2-macroglobulin. Biochem J. 1981 Jan 1;193(1):129–141. doi: 10.1042/bj1930129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Solomon E., Bobrow M., Goodfellow P. N., Bodmer W. F., Swallow D. M., Povey S., Noël B. Human gene mapping using an X/autosome translocation. Somatic Cell Genet. 1976 Mar;2(2):125–140. doi: 10.1007/BF01542626. [DOI] [PubMed] [Google Scholar]
  31. Solomon E., Swallow D., Burgess S., Evans L. Assignment of the human acid alpha-glucosidase gene (alphaGLU) to chromosome 17 using somatic cell hybrids. Ann Hum Genet. 1979 Jan;42(3):273–281. doi: 10.1111/j.1469-1809.1979.tb00661.x. [DOI] [PubMed] [Google Scholar]
  32. Sottrup-Jensen L., Hansen H. F., Mortensen S. B., Petersen T. E., Magnusson S. Sequence location of the reactive thiol ester in human alpha 2-macroglobulin. FEBS Lett. 1981 Jan 12;123(1):145–148. doi: 10.1016/0014-5793(81)80039-7. [DOI] [PubMed] [Google Scholar]
  33. Sottrup-Jensen L., Lønblad P. B., Stepanik T. M., Petersen T. E., Magnusson S., Jörnvall H. Primary structure of the 'bait' region for proteinases in alpha 2-macroglobulin. Nature of the complex. FEBS Lett. 1981 May 18;127(2):167–173. doi: 10.1016/0014-5793(81)80197-4. [DOI] [PubMed] [Google Scholar]
  34. Sottrup-Jensen L., Petersen T. E., Magnusson S. A thiol-ester in alpha 2-macroglobulin cleaved during proteinase complex formation. FEBS Lett. 1980 Dec 1;121(2):275–279. doi: 10.1016/0014-5793(80)80361-9. [DOI] [PubMed] [Google Scholar]
  35. Sottrup-Jensen L., Stepanik T. M., Kristensen T., Lønblad P. B., Jones C. M., Wierzbicki D. M., Magnusson S., Domdey H., Wetsel R. A., Lundwall A. Common evolutionary origin of alpha 2-macroglobulin and complement components C3 and C4. Proc Natl Acad Sci U S A. 1985 Jan;82(1):9–13. doi: 10.1073/pnas.82.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sottrup-Jensen L., Stepanik T. M., Wierzbicki D. M., Jones C. M., Lønblad P. B., Kristensen T., Mortensen S. B., Petersen T. E., Magnusson S. The primary structure of alpha 2-macroglobulin and localization of a Factor XIIIa cross-linking site. Ann N Y Acad Sci. 1983;421:41–60. doi: 10.1111/j.1749-6632.1983.tb18091.x. [DOI] [PubMed] [Google Scholar]
  37. Southern E. Gel electrophoresis of restriction fragments. Methods Enzymol. 1979;68:152–176. doi: 10.1016/0076-6879(79)68011-4. [DOI] [PubMed] [Google Scholar]
  38. Staden R. An interactive graphics program for comparing and aligning nucleic acid and amino acid sequences. Nucleic Acids Res. 1982 May 11;10(9):2951–2961. doi: 10.1093/nar/10.9.2951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Staden R. Automation of the computer handling of gel reading data produced by the shotgun method of DNA sequencing. Nucleic Acids Res. 1982 Aug 11;10(15):4731–4751. doi: 10.1093/nar/10.15.4731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Staden R. Graphic methods to determine the function of nucleic acid sequences. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):521–538. doi: 10.1093/nar/12.1part2.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Starkey P. M., Barrett A. J. Evolution of alpha 2-macroglobulin. The demonstration in a variety of vertebrate species of a protein resembling human alpha 2-macroglobulin. Biochem J. 1982 Jul 1;205(1):91–95. doi: 10.1042/bj2050091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Studencki A. B., Wallace R. B. Allele-specific hybridization using oligonucleotide probes of very high specific activity: discrimination of the human beta A- and beta S-globin genes. DNA. 1984;3(1):7–15. doi: 10.1089/dna.1.1984.3.7. [DOI] [PubMed] [Google Scholar]
  43. Swallow D. M., Solomon E., Pajunen L. Immunochemical analysis of the N-acetyl hexosaminidases in human-mouse hybrids made using a double selective system. Cytogenet Cell Genet. 1977;18(3):136–148. doi: 10.1159/000130758. [DOI] [PubMed] [Google Scholar]
  44. Swenson R. P., Howard J. B. Amino acid sequence of the tryptic peptide containing the alkylamine-reactive site from human alpha 2-macroglobulin. Identification of gamma-glutamylmethylamide. J Biol Chem. 1980 Sep 10;255(17):8087–8091. [PubMed] [Google Scholar]
  45. Tack B. F. The beta-Cys-gamma-Glu thiolester bond in human C3, C4, and alpha 2-macroglobulin. Springer Semin Immunopathol. 1983;6(4):259–282. doi: 10.1007/BF02116276. [DOI] [PubMed] [Google Scholar]
  46. Wang D., Yuan A. I., Feinman R. D. Structure of alpha 2-macroglobulin-protease complexes. Ann N Y Acad Sci. 1983;421:90–97. doi: 10.1111/j.1749-6632.1983.tb18095.x. [DOI] [PubMed] [Google Scholar]
  47. Waterfield M. D., Scrace G. T., Whittle N., Stroobant P., Johnsson A., Wasteson A., Westermark B., Heldin C. H., Huang J. S., Deuel T. F. Platelet-derived growth factor is structurally related to the putative transforming protein p28sis of simian sarcoma virus. Nature. 1983 Jul 7;304(5921):35–39. doi: 10.1038/304035a0. [DOI] [PubMed] [Google Scholar]
  48. Whitehead A. S., Solomon E., Chambers S., Bodmer W. F., Povey S., Fey G. Assignment of the structural gene for the third component of human complement to chromosome 19. Proc Natl Acad Sci U S A. 1982 Aug;79(16):5021–5025. doi: 10.1073/pnas.79.16.5021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. de Bruijn M. H., Fey G. H. Human complement component C3: cDNA coding sequence and derived primary structure. Proc Natl Acad Sci U S A. 1985 Feb;82(3):708–712. doi: 10.1073/pnas.82.3.708. [DOI] [PMC free article] [PubMed] [Google Scholar]

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