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Thromb Haemost 1987; 57(03): 356-360
DOI: 10.1055/s-0038-1651133
Original Article
Schattauer GmbH Stuttgart

The Primary Structure of the Inhibitor of Tissue Plasminogen Activator Found in the Seeds of Erythrina caffra

F J Joubert
1   The National Chemical Research Laboratory, Council for Scientific and Industrial Research, Pretoria, Republic of South Africa
,
E B D Dowdle
2   The Department of Clinical Science and Immunology, University of Cape Town and The South African Medical Research Council Human Cell Biology Research Unit, Observatory, Republic of South Africa
› Author Affiliations
Further Information

Publication History

Received 05 December 1986

Accepted after revision 10 March 1987

Publication Date:
06 July 2018 (online)

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Summary

Trypsin and tissue plasminogen activator inhibitor DE-3 from Erythrina caffra contains 172 amino acids, including 4 half-cystine residues, and resembles the Kunitz-type inhibitors. Limited hydrolysis of DE-3 with trypsin at pH 3.2 produced two fragments, FI and F2, containing 63 and 109 amino acids, respectively. Amino-terminal sequence studies showed that FI was the N-terminal and that F2 was the C-terminal fragment. The complete amino acid sequence of the fragments were then determined on peptides produced by enzymatic digestion with trypsin. The sequence of trypsin and tissue plasminogen activator inhibitor DE-3 from E. caffra seeds shows a high degree of homology to that of trypsin and tissue plasminogen activator inhibitor DE-3 from E. latissima seeds and revealed only four amino acids which were replaced.

Keywords

Erythrina caffra - Trypsin and tissue plasminogen activator inhibitor - Amino acid sequence
 

  • References

  • 1 Kunitz M. Crystalline soybean trypsin inhibitor II. General properties. J Gen Physiol 1947; 30: 291-310
    CrossrefPubMedGoogle Scholar
  • 2 Kunitz M. Isolation of a crystalline protein compound of trypsin and of soybean trypsin-inhibitor. J Gen Physiol 1947; 30: 311-320
    CrossrefPubMedGoogle Scholar
  • 3 Yamamoto M, Hara S, Ikenaka T. Amino acid sequences of two trypsin inhibitors from Winged bean seeds (Psophocarpus tetragonolobus [L] DC). J Biochem 1983; 94: 849-863
    CrossrefPubMedGoogle Scholar
  • 4 Laskowski Jr M, Kato I. Protein inhibitors of proteinases. Ann Rev Biochem 1980; 49: 593-626
    CrossrefPubMedGoogle Scholar
  • 5 Joubert FJ, Carlsson F HH, Haylett T. Purification of two proteinase inhibitors (DE-1 and DE-3) from Erythrina latissima (Broad-leaved Erythrina) seed. Hoppe-Seyler’s Z Physiol Chem 1981; 362: 531-538
    PubMedGoogle Scholar
  • 6 Joubert FJ, Heussen C, Dowdle E BD. The complete amino acid sequence of trypsin inhibitor DE-3 from Erythrina latissima seeds. J Biol Chem 1985; 260: 12948-12953
    PubMedGoogle Scholar
  • 7 Koide T, Ikenaka T. Studies on soybean trypsin inhibitors. 1. Fragmentation of soybean trypsin inhibitor (Kunitz) by limited proteolysis and by chemical cleavage. Eur J Biochem 1973; 32: 401-407
    CrossrefPubMedGoogle Scholar
  • 8 Koide T, Ikenaka T. Studies on soybean trypsin inhibitors. 3. Amino acid sequence of the carboxy-terminal region and the complete sequence of soybean trypsin inhibitor (Kunitz). Eur J Biochem 1973; 32: 417-431
    CrossrefPubMedGoogle Scholar
  • 9 Koide T, Tsunasawa S, Ikenaka T. Studies on soybean trypsin inhibitors. 2. Amino acid sequence around the reactive site of soybean trypsin inhibitor (Kunitz). Eur J Biochem 1973; 32: 408-416
    CrossrefPubMedGoogle Scholar
  • 10 Heussen C, Joubert F, Dowdle E BD. Purification of human tissue plasminogen activator with Erythrina trypsin inhibitor. J Biol Chem 1984; 259: 11635-11638
    PubMedGoogle Scholar
  • 11 Barneby RC, Krukoff BA, Raven PH. Erythrina Symposium. IV Erythrina (Fabaceae: Faboideae). Allertonia Pacific Tropical Botanical Garden, Lawai, Kauai, Hawaii 1982; 3: 154-165
    PubMedGoogle Scholar
  • 12 Joubert FJ, Merrifield EH, Dowdle E BD. The reactive sites of proteinase inhibitors from Erythrina seeds. Int J Biochem. (submitted for publication)
    PubMedGoogle Scholar
  • 13 Joubert FJ. Purification and properties of the proteinase inhibitors from Erythrina caffra (coast Erythrina) seed. Int J Biochem 1982; 14: 187-193
    CrossrefPubMedGoogle Scholar
  • 14 Brandt WF, Alk H, Chauhan Hauhan M, von Holt C. A simple modification converts the spinning cup protein sequencer into a vapour-phase sequencer. FEBS Lett 1984; 174: 228-232
    CrossrefPubMedGoogle Scholar
  • 15 Hewick RM, Hunkapiller MW, Hood LE, Dreyer WJ. A gas-liquid solid phase peptide and protein sequenator. J Biol Chem 1981; 256: 7990-7997
    PubMedGoogle Scholar
  • 16 Wolfenden R, Andersson L, Cullis PM, Southgate C CB. Affinities of amino acid side chains for solvent water. Biochemistry 1981; 20: 849-855
    CrossrefPubMedGoogle Scholar
  • 17 Tanford C. The interpretation of hydrogen ion titration. Curves of patients. Adv Protein Chem 1962; 17: 69-165
    PubMedGoogle Scholar
  • 18 Bergmann SR, Fox K AA, Ter-Pogossian MM, Sobel BE, Collen D. Clot-selective coronary thrombolysis with tissue-type plasminogen activator. Science 1983; 220: 1181-1183
    CrossrefPubMedGoogle Scholar
  • 19 Van de Werf F, Bergmann SR, Fox K AA, De Geest H, Hoyng CF, Sobel BE, Collen D. Coronary thrombolysis with intravenously administered human tissue-type plasminogen activator produced by recombinant DNA technology. Circulation 1984; 69: 605-610
    CrossrefPubMedGoogle Scholar