1887

Abstract

A method comprising enzyme separation by SDS-PAGE and subsequent use of peptidyl aminomethylcoumarins as substrates has been used to study proteinases of the protozoan parasite . The application of this method has allowed investigation of the substrate specificities of individual proteinases in cell lysates without the need for enzyme purification. The results show that contains a group of cysteine proteinases, probably four in number, with substrate and inhibitor specificities similar to those of cathepsin L. A second group of proteinases, larger enzymes with significantly different substrate specificities and sensitivity to inhibitors, was also detected. Peptidyl diazomethanes inhibited the cysteine proteinases and also parasite growth, offering promise that peculiarities in the substrate specificity of trypanosomal cysteine proteinases could be exploited by compounds of this type.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-136-5-921
1990-05-01
2021-10-27
Loading full text...

Full text loading...

/deliver/fulltext/micro/136/5/mic-136-5-921.html?itemId=/content/journal/micro/10.1099/00221287-136-5-921&mimeType=html&fmt=ahah

References

  1. Barrett A.J., Buttle D.J., Mason R.W. 1988; Lysosomal cysteine proteinases. ISI Atlas of Science: Biochemistry 1256–260
    [Google Scholar]
  2. Brocklehurst K. 1987; Acyl group transfer - cysteine proteinases. In Enzyme Mechanisms pp. 140–158 Page M.I., Williams A. Edited by London: Royal Society of Chemistry;
    [Google Scholar]
  3. Brun R., Schonenberger M. 1979; Cultivation and in vitro cloning of procyclic culture forms of Trypanosoma hrucei in semi- defined medium. Acta Tropica 36:289–292
    [Google Scholar]
  4. Crawford C., Mason R.W., Wikstrom P., Shaw E. 1988; The design of peptidyldiazomethane inhibitors to distinguish between the cysteine proteinases calpain II, cathepsin L and cathepsin B. Biochemical Journal 253:751–758
    [Google Scholar]
  5. Koehler S., Ho -H.D. 1988; Purification and characterization of gibberellic acid-induced cysteine endoproteases in barley aleurone layers. Plant Physiology 87:95–103
    [Google Scholar]
  6. Lanham S.N., Godfrey D.G. 1970; Isolation of salivarian trypanosomes from man and other mammals using DEAE cellulose. Experimental Parasitology 20:521–534
    [Google Scholar]
  7. Lockwood B.C., North M.J., Mallinson D.J., Coombs G.H. 1987; Analysis of Leishmania proteinases reveals developmental changes in species-specific forms and a common 68 kDa activity. FEMS Microbiology Letters 48:345–350
    [Google Scholar]
  8. Lonsdale-Eccles J.D., Grab D.J. 1987; Lysosomal and nonlysosomal peptidyl hydrolases of the bloodstream forms of Trypanosoma brucei brucei. European Journal of Biochemistry 169:467–475
    [Google Scholar]
  9. Lonsdale-Eccles J.D., Mpimbaza G.W.N. 1986; Thiol-dependent proteases of African trypanosomes. European Journal of Biochemistry 155:469–473
    [Google Scholar]
  10. Mottram J.C., North M.J., Barry J.D., Coombs G.H. 1989; A cysteine proteinase cDNA from Trypanosoma brucei predicts an enzyme with an unusual C-terminal extension. FEBS Letters 258:211–215
    [Google Scholar]
  11. North M.J., Coombs G.H., Barry J.D. 1983; A comparative study of the proteolytic enzymes of Trypanosoma brucei, T. equiperdum, T. evansi, T. vivax, Leishmania tarentolae and Crithidia fasciculata. Molecular and Biochemical Parasitology 9:161–180
    [Google Scholar]
  12. North M.J., Robertson C.D., Coombs G.H. 1990; The specificity of trichomonad cysteine proteinases analysed using fluorogenic substrates and specific inhibitors. Molecular and Biochemical Parasitology 39:183–194
    [Google Scholar]
  13. Pamer E.G., So M., Davis C.E. 1989; Identification of a developmentally regulated cysteine protease of Trypanosoma brucei. Molecular and Biochemical Parasitology 33:27–32
    [Google Scholar]
  14. Sakanari J.A., Staunton C.E., Eakin A.E., Craik C.S., Mckerrow J.H. 1989; Serine proteases from nematode and protozoan parasites: isolation of sequence homologs using generic molecular probes. Proceedings of the National Academy of Sciences of the United States of America 86:4863–4867
    [Google Scholar]
  15. Schechter I., Berger A. 1967; On the size of the active site in proteases. I. Papain. Biochemical and Biophysical Research Communications 27:157–162
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-136-5-921
Loading
/content/journal/micro/10.1099/00221287-136-5-921
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error