Kinetic properties of triose-phosphate isomerase from Trypanosoma brucei brucei. A comparison with the rabbit muscle and yeast enzymes.

The kinetic properties of Trypanosoma brucei brucei triose-phosphate isomerase are compared with those of the commercially available rabbit muscle and yeast enzymes and with published data on the chicken muscle enzyme. With glyceraldehyde 3-phosphate as substrate Km = 0.25 +/- 0.05 mM and kcat = 3.7 X 10(5) min-1. With dihydroxyacetone phosphate as substrate Km = 1.2 +/- 0.1 mM and kcat = 6.5 X 10(4) min-1. The pH dependence of Km and Vmax at 0.1 M ionic strength is in agreement with the results published for the yeast and chicken muscle enzymes. At ionic strength below 0.05 M the effect of a charged group specific for the trypanosomal enzyme and absent from the yeast and rabbit muscle enzymes becomes detectable. This effect significantly increases Km whereas Vmax becomes slightly higher. Trypanosomal triose-phosphate isomerase is inhibited by sulphate, phosphate and arsenate ions, by 2-phosphoglycolate and a number of documented inhibitors in the same concentration range as are the other triose-phosphate isomerases. The trypanocidal drug, Suramin inhibits T. brucei and rabbit muscle triose-phosphate isomerase to the same extent while leaving the yeast enzyme relatively unaffected.

[1]  M. Dixon.  The determination of enzyme inhibitor constants. , 1953, The Biochemical journal.

[2]  Farrington Daniels,et al.  Physical Chemistry, 2nd Ed. , 1961 .

[3]  E. Noltmann,et al.  Reaction of Sulfhydryl Groups in Phosphoglucose Isomerase with Organic Mercurials , 1967 .

[4]  T. Bücher,et al.  The isolation and crystallization of yeast and rabbit liver triose phosphate isomerase and a comparative characterization with the rabbit muscle enzyme. , 1970, European journal of biochemistry.

[5]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[6]  J. Knowles,et al.  pH-dependence of the triose phosphate isomerase reaction. , 1972, The Biochemical journal.

[7]  A. Coulson,et al.  Specificity and kinetics of triose phosphate isomerase from chicken muscle. , 1972, The Biochemical journal.

[8]  S. Waley,et al.  Refolding of triose phosphate isomerase. , 1973, The Biochemical journal.

[9]  J. Harris,et al.  Triose phosphate isomerase from the coelacanth. An approach to the rapid determination of an amino acid sequence with small amounts of material. , 1974, The Biochemical journal.

[10]  D. D. Perrin,et al.  Buffers for pH and metal ion control , 1974 .

[11]  G. Petsko,et al.  Structure of chicken muscle triose phosphate isomerase determined crystallographically at 2.5Å resolution: using amino acid sequence data , 1975, Nature.

[12]  F. C. Hartman,et al.  The influence of pH on the interaction of inhibitors with triosephosphate isomerase and determination of the pKa of the active-site carboxyl group. , 1975, Biochemistry.

[13]  L. Jaenicke,et al.  Buffers for pH and Metal Ion Control. Von D. D. Perrin und B. Dempsey. Chapman and Hall, London 1974. 1. Aufl., VII, 176 S., zahlr. Tab., geb. £ 3.50 , 1975 .

[14]  P. Corran,et al.  The amino acid sequence of rabbit muscle triose phosphate isomerase. , 1975, The Biochemical journal.

[15]  F. C. Hartman,et al.  [95] Triosephosphate isomerase from rabbit muscle , 1975 .

[16]  J. Knowles,et al.  Free-energy profile of the reaction catalyzed by triosephosphate isomerase. , 1976, Biochemistry.

[17]  F. Opperdoes,et al.  Localization of nine glycolytic enzymes in a microbody‐like organelle in Trypanosoma brucei: The glycosome , 1977, FEBS letters.

[18]  F. C. Hartman,et al.  Apparent equivalence of the active-site glutamyl residue and the essential group with pKalpha 6.0 in triosephosphate isomerase. , 1977, Biochemical and biophysical research communications.

[19]  J. Knowles,et al.  Critical ionization states in the reaction catalyzed by triosephosphate isomerase. , 1978, Biochemistry.

[20]  J. Harris,et al.  Primary structure of triosephosphate isomerase from Bacillus stearothermophilus. , 1980, European journal of biochemistry.

[21]  D W Banner,et al.  On the three-dimensional structure and catalytic mechanism of triose phosphate isomerase. , 1981, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[22]  F. C. Hartman,et al.  Crystallization of yeast triose phosphate isomerase from polyethylene glycol. Protein crystal formation following phase separation. , 1981, The Journal of biological chemistry.

[23]  R. Wierenga,et al.  Preliminary crystallographic studies of triosephosphate isomerase from the blood parasite Trypanosoma brucei brucei. , 1984, Journal of molecular biology.

[24]  I. Coppens,et al.  Purification, morphometric analysis, and characterization of the glycosomes (microbodies) of the protozoan hemoflagellate Trypanosoma brucei , 1984, The Journal of cell biology.

[25]  F. Opperdoes,et al.  Simultaneous purification of hexokinase, class-I fructose-bisphosphate aldolase, triosephosphate isomerase and phosphoglycerate kinase from Trypanosoma brucei. , 1984, European journal of biochemistry.

[26]  R. Howells.  The modes of action of some anti-protozoal drugs , 1985, Parasitology.

[27]  F. Opperdoes.  Biochemical peculiarities of trypanosomes, African and South American. , 1985, British medical bulletin.

[28]  P. Borst,et al.  Characterization of the gene for the microbody (glycosomal) triosephosphate isomerase of Trypanosoma brucei. , 1986, The EMBO journal.

[29]  F. Opperdoes,et al.  Glycolytic enzymes of Trypanosoma brucei. Simultaneous purification, intraglycosomal concentrations and physical properties. , 1986, European journal of biochemistry.

[30]  F. Opperdoes,et al.  The phosphoglycerate kinases from Trypanosoma brucei. A comparison of the glycosomal and the cytosolic isoenzymes and their sensitivity towards suramin. , 1987, European journal of biochemistry.

[31]  F. Opperdoes,et al.  Common elements on the surface of glycolytic enzymes from Trypanosoma brucei may serve as topogenic signals for import into glycosomes. , 1987, The EMBO journal.