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. 1994 Mar 15;298(Pt 3):619–622. doi: 10.1042/bj2980619

Production of human embryonic haemoglobin (Gower II) in a yeast expression system.

R M Mould 1, O M Hofmann 1, T Brittain 1
PMCID: PMC1137904  PMID: 8141775

Abstract

The cDNA coding for a human embryonic globin protein has been obtained from an erythroleukaemic cell line. A plasmid expression system for human embryonic haemoglobin Gower II containing cDNA copies of the appropriate pair of globin genes coupled to synthetic galactose-regulated hybrid promoters has been engineered. Transformation of Saccharomyces cerevisiae with this plasmid yields a cellular system capable of high-level production of fully functional tetrameric embryonic haemoglobin. We have developed a purification scheme which gives high yields of pure human embryonic haemoglobin suitable for structural and functional studies. Preliminary characterization studies are reported.

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Selected References

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  1. Adachi K., Konitzer P., Lai C. H., Kim J., Surrey S. Oxygen binding and other physical properties of human hemoglobin made in yeast. Protein Eng. 1992 Dec;5(8):807–810. doi: 10.1093/protein/5.8.807. [DOI] [PubMed] [Google Scholar]
  2. Andersson L. C., Nilsson K., Gahmberg C. G. K562--a human erythroleukemic cell line. Int J Cancer. 1979 Feb;23(2):143–147. doi: 10.1002/ijc.2910230202. [DOI] [PubMed] [Google Scholar]
  3. Brittain T. A possible role for protein dissociation in the functioning of embryonic haemoglobins. J Exp Biol. 1987 Jan;127:443–447. doi: 10.1242/jeb.127.1.443. [DOI] [PubMed] [Google Scholar]
  4. Gale R. E., Clegg J. B., Huehns E. R. Human embryonic haemoglobins Gower 1 and Gower 2. Nature. 1979 Jul 12;280(5718):162–164. doi: 10.1038/280162a0. [DOI] [PubMed] [Google Scholar]
  5. Hayashi A., Suzuki T., Shin M. An enzymic reduction system for metmyoglobin and methemoglobin, and its application to functional studies of oxygen carriers. Biochim Biophys Acta. 1973 Jun 15;310(2):309–316. doi: 10.1016/0005-2795(73)90110-4. [DOI] [PubMed] [Google Scholar]
  6. Hill J., Donald K. A., Griffiths D. E., Donald G. DMSO-enhanced whole cell yeast transformation. Nucleic Acids Res. 1991 Oct 25;19(20):5791–5791. doi: 10.1093/nar/19.20.5791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hoffman S. J., Looker D. L., Roehrich J. M., Cozart P. E., Durfee S. L., Tedesco J. L., Stetler G. L. Expression of fully functional tetrameric human hemoglobin in Escherichia coli. Proc Natl Acad Sci U S A. 1990 Nov;87(21):8521–8525. doi: 10.1073/pnas.87.21.8521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Huehns E. R., Faroqui A. M. Oxygen dissociation properties of human embryonic red cells. Nature. 1975 Mar 27;254(5498):335–337. doi: 10.1038/254335a0. [DOI] [PubMed] [Google Scholar]
  9. Lozzio C. B., Lozzio B. B. Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome. Blood. 1975 Mar;45(3):321–334. [PubMed] [Google Scholar]
  10. Martin de Llano J. J., Schneewind O., Stetler G., Manning J. M. Recombinant human sickle hemoglobin expressed in yeast. Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):918–922. doi: 10.1073/pnas.90.3.918. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mattia E., Josic D., Ashwell G., Klausner R., van Renswoude J. Regulation of intracellular iron distribution in K562 human erythroleukemia cells. J Biol Chem. 1986 Apr 5;261(10):4587–4593. [PubMed] [Google Scholar]
  12. McBride L. J., Koepf S. M., Gibbs R. A., Salser W., Mayrand P. E., Hunkapiller M. W., Kronick M. N. Automated DNA sequencing methods involving polymerase chain reaction. Clin Chem. 1989 Nov;35(11):2196–2201. [PubMed] [Google Scholar]
  13. Popp R. A., Marsh C. L., Skow L. C. Expression of embryonic hemoglobin genes in mice heterozygous for alpha-thalassemia or beta-duplication traits and in mice heterozygous for both traits. Dev Biol. 1981 Jul 15;85(1):123–128. doi: 10.1016/0012-1606(81)90241-4. [DOI] [PubMed] [Google Scholar]
  14. Purdie A., Wells R. M., Brittain T. Molecular aspects of embryonic mouse haemoglobin ontogeny. Biochem J. 1983 Nov 1;215(2):377–383. doi: 10.1042/bj2150377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rutherford T. R., Clegg J. B., Weatherall D. J. K562 human leukaemic cells synthesise embryonic haemoglobin in response to haemin. Nature. 1979 Jul 12;280(5718):164–165. doi: 10.1038/280164a0. [DOI] [PubMed] [Google Scholar]
  16. Testa U., Vainchenker W., Beuzard Y., Rouyer-Fessard P., Guerrasio A., Titeux M., Lapotre P., Bouguet J., Breton-Gorius J., Rosa J. Hemoglobin expression in clones of K562 cell line. Eur J Biochem. 1982 Jan;121(3):649–655. doi: 10.1111/j.1432-1033.1982.tb05835.x. [DOI] [PubMed] [Google Scholar]
  17. Tuchinda S., Nagai K., Lehmann H. Oxygen dissociation curve of haemoglobin Portland. FEBS Lett. 1975 Jan 1;49(3):390–391. doi: 10.1016/0014-5793(75)80792-7. [DOI] [PubMed] [Google Scholar]
  18. Tyuma I., Shimizu K. Effect of organic phosphates on the difference in oxygen affinity between fetal and adult human hemoglobin. Fed Proc. 1970 May-Jun;29(3):1112–1114. [PubMed] [Google Scholar]
  19. Villeponteau B., Martinson H. Isolation and characterization of the complete chicken beta-globin gene region: frequent deletion of the adult beta-globin genes in lambda. Nucleic Acids Res. 1981 Aug 11;9(15):3731–3746. doi: 10.1093/nar/9.15.3731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wagenbach M., O'Rourke K., Vitez L., Wieczorek A., Hoffman S., Durfee S., Tedesco J., Stetler G. Synthesis of wild type and mutant human hemoglobins in Saccharomyces cerevisiae. Biotechnology (N Y) 1991 Jan;9(1):57–61. doi: 10.1038/nbt0191-57. [DOI] [PubMed] [Google Scholar]
  21. Weber R. E., Kleinschmidt T., Braunitzer G. Embryonic pig hemoglobins Gower I (zeta 2 epsilon 2), Gower II (alpha 2 epsilon 2), Heide I (zeta 2 theta 2) and Heide II (alpha 2 theta 2): oxygen-binding functions related to structure and embryonic oxygen supply. Respir Physiol. 1987 Sep;69(3):347–357. doi: 10.1016/0034-5687(87)90088-0. [DOI] [PubMed] [Google Scholar]
  22. Wood W. G., Clegg J. B., Weatherall D. J. Developmental biology of human hemoglobins. Prog Hematol. 1977;10:43–90. [PubMed] [Google Scholar]

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