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Volume 144, Issue 4

Molecular analysis of Physarum haemagglutinin l: lack of a signal sequence, sulphur amino acids and post-translational modifications Free

Abstract

The cDNAs encoding haemagglutinin I from plasmodia of have been cloned using PCR protocols. The composite haemagglutinin I cDNA sequence, derived from several overlapping clones from PCR fragments, spans 408 nt and the 315 bp ORF encodes a polypeptide of 104 aa without a typical signal sequence. The putative molecular mass deduced from the amino acid sequence (10760.76 Da) corresponds exactly to that determined by electrospray ionization MS (10759.86±.15 Da), suggesting that haemagglutinin I is not subject to post-translational modification. Haemagglutinin I lacks sulphur amino acids and has a β-sheet as the major secondary structure. Expression of the coding sequence in yielded a product that exhibits the same sugar-binding specificity as natural haemagglutinin I. The deduced amino acid sequence shows little similarity to that of any known lectins and thus apparently represents a novel type of lectin.

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Keyword(s): cDNA cloning , haemagglutinin , lectin , Physarum polycephalum and post-translational modification
© Society for General Microbiology 1998
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1998-04-01
2025-04-19
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References

  1. Avichezer D., Katcoff D. J., Garber N. C., Gilboa-Garber N. (1992); Analysis of the amino acid sequence of the Pseudomonas aeruginosa galactophilic PA-I lectin.. Journal of Biological Chemistry, 267:(32),23023–23027 [View Article]
     [Google Scholar]
  2. Avichezer D., Gilboa-Garber N., Garber N.C., Katcoff D.J. (1994); Pseudomonas aeruginosa PA-I lectin gene molecular analysis and expression in Escherichia coli.. Biochimica Et Biophysica Acta, 1218:(1),11–20 [View Article]
     [Google Scholar]
  3. Barondes S. H., Springer W. R. (1987) An endogenous lectin and an oligosaccharide participate in adhesion mechanisms in Dictyostelium.. Edited by Loomis W. F. In Genetic Regulation of Development New York:: Alan R. Liss;129–140
     [Google Scholar]
  4. Barondes S. H., Cooper D. N. W., Gitt M. A., Leffler H. (1994); Galectins. Structure and function of a large family of animal lectins.. Journal of Biological Chemistry, 269:(33),20807–20810 [View Article]
     [Google Scholar]
  5. Chou P. Y., Fasman G. D. (1978); Prediction of the secondary structure of proteins from their amino acid sequence.. Adv Enzymol, 47:45–148
     [Google Scholar]
  6. Cooper D. N. W., Lee S.-C., Barondes S. H. (1983); Discoidin- binding polysaccharide from Dictyostelium discoideum.. Journal of Biological Chemistry, 258:(14),8745–8750 [View Article]
     [Google Scholar]
  7. Cooper D. N. W., Boulianne R. P., Charlton S., Farrell E. M., Sucher A. et al. (1997); Fungal galectins, sequence and specificity of two isolectins from Coprinus cinereus.. ] Biol Chem, 272:(3),1514–1521 [View Article]
     [Google Scholar]
  8. Daniel J. W., Baldwin H. H. (1964); Axenic culture of Physarum polycephalum on soluble media.. Methods Cell Physiol, 1:15–29
     [Google Scholar]
  9. Frohman M. A., Dush M. K., Martin G. R. (1988); Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer.. Proc Natl Acad Sci USA, 85:(23),8998–9002 [View Article]
     [Google Scholar]
  10. Gowda L. R., Savithri H. S., Rao D. R. (1994); The complete primary structure of a unique mannose/glucose-specific lectin from Field Bean (Dolichos Lab lab).. Journal of Biological Chemistry, 269:1878918793
     [Google Scholar]
  11. Hirabayashi J., Kasai K. (1993); The family of metazoan metalindependent beta-galactoside-binding lectins: structure, function and molecular evolution.. Glycobiology, 3:(4),297–304 [View Article]
     [Google Scholar]
  12. Inbar J., Chet I. (1994); A newly isolated lectin from the plant pathogenic fungus Sclerotium rolfsii: purification, characterization and role in mycoparasitism.. Microbiology, 140:(3),651–657 [View Article]
     [Google Scholar]
  13. Kyte J. St Doolittle, R. F. (1982); A simple method for displaying the hydropathic character of a protein.. Journal of Molecular Biology, 157:(1),105–132 [View Article]
     [Google Scholar]
  14. Lobsanov Y. D., Gitt M. A., Leffler H., Barondes S. H., Rini J. M. (1993); X-ray crystal structure of the human dimeric S-Lac lectin, L-14-II, in complex with lactose at 2-9-A resolution.. Journal of Biological Chemistry, 268:(36),27034–27038 [View Article]
     [Google Scholar]
  15. Matsudaira P. (1987); Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes.. Journal of Biological Chemistry, 262:(21),10035–10038 [View Article]
     [Google Scholar]
  16. Morita M., Nishi A. (1991); Glycoproteins and enzymes associated with plasmodial membrane and slime layer of Physarum polycephalum.. Journal of General and Applied Microbiology, 37:(1),93–109 [View Article]
     [Google Scholar]
  17. Morita M., Nishi A. (1992); Isolation and partial characterization of haemagglutinins from plasmodia of Physarum polycephalum.. Journal of General Microbiology, 138:(3),619–625 [View Article]
     [Google Scholar]
  18. Morita M., Minami E., Zheng L., Nishi A. (1995); Physarum polycephalum haemagglutinins: effect of nutrition on synthesis, and their possible role in nature.. Microbiology, 141:(9),2315–2322 [View Article]
     [Google Scholar]
  19. Muramoto K., Kamiya H. (1992); The amino-acid sequence of a lectin from conger eel, Conger myriaster, skin mucus.. Biochimica Et Biophysica Acta, 1116:(2),129–136 [View Article]
     [Google Scholar]
  20. Nishimura M., Tanigaki C, Okamura S. (1991); Changes in fi- tubulin isoforms and their RNA level in synchronized tobacco cells.. Cell Struct Punct, 16:(6),489–494 [View Article]
     [Google Scholar]
  21. Ozeki Y., Matsui T., Suzuki M., Titani K. (1991); Amino acid sequence and molecular characterization of a D-galactoside- specific lectin purified from sea urchin (Anthocidaris crassispina} eggs.. Biochemistry, 30:(9),2391–2394 [View Article]
     [Google Scholar]
  22. Reiman D. A., Domenighini M., Tomanen E., Rappuoli R., Falkow S. (1989); Filamentous hemagglutinin of Bordetella pertussis: nucleotide sequence and crucial role in adherence.. Proc Natl Acad Sci USA, 86:(8),2637–2641 [View Article]
     [Google Scholar]
  23. Rosen S., Ek L., Rask L., Tunlid A. (1992); Purification and characterization of a surface lectin from the nematode-trapping fungus Arthrobotrys oligospora.. Journal of General Microbiology, 138:(12),2663–2672 [View Article]
     [Google Scholar]
  24. Rosen S., Bergstrom J., Karlsson K.-A., Tunlid A. (1996a); A multispecific saline-soluble lectin from the parasitic fungus Arthrobotrys oligospora. Similarities in the binding specificities compared with a lectin from the mushroom Agaricus bisporus.. European Journal of Biochemistry, 238:(3),830–837 [View Article]
     [Google Scholar]
  25. Rosen S., Kata M., Persson Y., Lipniunas P. H., Wikstrom M., van den Hondel C. A. M. J. J, van den Brink J.M., Rask L., Heden L.-0., Tunlid A. (1996b); Molecular characterization of a saline-soluble lectin from a parasitic fungus. Extensive sequence similarities between fungal lectins.. European Journal of Biochemistry, 238:(3),822–829 [View Article]
     [Google Scholar]
  26. Sambrook J., Fritsch E. F., Maniatis T. (1989) Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory;
     [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. (1977); DNA sequencing with chain-terminating inhibitors.. Proc Natl Acad Sci USA, 74:(12),5463–5467 [View Article]
     [Google Scholar]
  28. Schagger H., van Jagow G. (1987); Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa.. Analytical Biochemistry, 166:(2),368–379 [View Article]
     [Google Scholar]
  29. Schreckenbach T., Werenskiold A.-K. (1986) Gene expression during plasmodial differentiation.. Edited by Dove W. F., Dee J., Hatano S., Haugli F. B., Wohlfarth-Bottermann K.-E. In The Molecular Biology of Physarum polycephalum New York:: Plenum Press;131–150
     [Google Scholar]
  30. Sharon N. (1993); Lectin-carbohydrate complexes of plants and animals: an atomic view.. Trends in Biochemical Sciences, 18:(6),221–226 [View Article]
     [Google Scholar]
  31. Van Damme E. J. M., Barre A., Verhaert P., Rouge P., Peumans W. J. (1996); Molecular cloning of the mitogenic mannose/maltose-specific rhizome lectin from Calystegia sepium.. Febs Letters, 397:(2–3),352–356 [View Article]
     [Google Scholar]
  32. Vranken A. M., Van Damme E. J. M., Allen A. K., Peumans W. J. (1987); Purification and properties of an N-acetylgalactosamine specific lectin from the plant pathogenic fungus Rhizoctonia solani.. Febs Letters, 216:(1),67–72 [View Article]
     [Google Scholar]
  33. Yatohgo T., Nakata M., Tumuraya Y., Hashimoto Y., Yamamoto S. (1988); Purification and properties of a lectin from the fruitbodies of Flammulina velutipes.. Agric Biol Chem, 52:1485–1493
     [Google Scholar]
  34. Young N. M., Johnston R. A. Z., Szabo A. G., Watson D. C. (1989); Homology of the D-galactose-specific lectins from Arto- carpus integrifolia and Maclura pomifera and the role of an unusual small polypeptide subunit.. Archives of Biochemistry and Biophysics, 270:(2),596–603 [View Article]
     [Google Scholar]
  35. Young N. M., Watson D. C., Yaguchi M., Ader R., Arango R. et al. (1995); C-Terminal post-translational proteolysis of plant lectins and their recombinant forms expressed in Escherichia coli.. ] Biol Chem, 270:(6),2563–2570 [View Article]
     [Google Scholar]
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Molecular analysis of Physarum haemagglutinin l: lack of a signal sequence, sulphur amino acids and post-translational modifications
Microbiology 144, 1077 (1998); https://doi.org/10.1099/00221287-144-4-1077
/content/journal/micro/10.1099/00221287-144-4-1077
/content/journal/micro/10.1099/00221287-144-4-1077
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