Selective Interaction of Glycosomal Enzymes from Trypanosoma brucei with Hydrophobic Cyclic Hexapeptides

Callens, Mia; Vanroy, J.; Zeelen, J.P.; Borchert, Torben V.; Nalis, Dominique; Wierenga, R.K.; Opperdoes, Frederik

doi:10.1006/bbrc.1993.2097

Cited by 4 publications

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“…Stabilization of this reverse turn feature in further analogs merits investigation. Interestingly, sequence‐unrelated cyclic hexapeptides have been shown to be effective inhibitors of trypanosomal TIM [42–44]. The present study demonstrates that the use of dimer interface peptides is a viable strategy in generating lead sequences for inhibitor design.…”

Section: Discussionmentioning

confidence: 70%

Synthetic peptides as inactivators of multimeric enzymes: inhibition of Plasmodium falciparum triosephosphate isomerase by interface peptides

et al. 2001

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Synthetic peptides corresponding to two distinct segments of the subunit interface of the homodimeric enzyme triosephosphate isomerase (residues 9^18, ANWKCNGTLE, peptide I; residues 68^79, KFGNGSYTGEVS, peptide II) from Plasmodium falciparum (PfTIM) have been investigated for their ability to act as inhibitors by interfering with the quaternary structure of the enzyme. An analog of peptide II containing cysteine at the site corresponding to position 74 and tyrosine at position 69 in the protein sequence KYGNGSCTGEVS (peptide III) was also investigated. A substantial fall in enzyme activity was observed following incubation of the enzyme with peptide II, whereas peptide I did not show any appreciable inhibition. The inhibitory effect was more pronounced on two mutants of PfTIM (Y74C and Y74G), both of which have reduced stability compared to the wild-type protein due to an interface cavity. The IC 50 value determined for peptide II is in the range of 0.60 .8 W WM. This study suggests that interface peptides of oligomeric enzymes can be used to inhibit dimeric enzymes by disrupting their native multimeric states and may provide lead structures for potential inhibitor design. ß 2001 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

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Section: Discussionmentioning

confidence: 70%

Synthetic peptides as inactivators of multimeric enzymes: inhibition of Plasmodium falciparum triosephosphate isomerase by interface peptides

et al. 2001

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“…This unusually high pI suggests that it might be possible to develop inhibitors selective for trypanosomal TIM as therapeutic reagents. Consequently, there have been extensive mechanistic and crystallographic ,− studies of trypanosomal TIM in an effort to gain insight that may help in guiding the design of Tbb TIM-specific tight-binding enzyme inhibitors. ,− …”

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confidence: 99%

Wildtype and Engineered Monomeric Triosephosphate Isomerase from Trypanosoma brucei: Partitioning of Reaction Intermediates in D₂O and Activation by Phosphite Dianion

et al. 2011

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Product yields for the reactions of (R)-glyceraldehyde 3-phosphate (GAP) in D2O at pD 7.9 catalyzed by wildtype triosephosphate isomerase from Trypanosoma brucei brucei (Tbb TIM) and a monomeric variant (monoTIM) of this wildtype enzyme were determined by 1H NMR spectroscopy, and were compared with the yields determined in earlier work for the reactions catalyzed by TIM from rabbit and chicken muscle [O’Donoghue, A. C, Amyes, T. L. and Richard J.P. (2005), Biochemistry 44, 2610–2621]. Three products were observed from the reactions catalyzed by TIM: dihydroxyacetone phosphate (DHAP) from isomerization with intramolecular transfer of hydrogen, d-DHAP from isomerization with incorporation of deuterium from D2O into C-1 of DHAP, and d-GAP from incorporation of deuterium from D2O into C-2 of GAP. The yield of DHAP formed by intramolecular transfer of hydrogen decreases from 49% for the muscle enzymes to 40% for wildtype Tbb TIM to 34% for monoTIM. There is no significant difference in the ratio of the yields of d-DHAP and d-GAP for wildtype TIM from muscle sources and Trypanosoma brucei brucei, but partitioning of the enediolate intermediate of the monoTIM reaction to form d-DHAP is less favorable ((kC1)D/(kC2)D = 1.1) than for the wildtype enzyme ((kC1)D/(kC2)D = 1.7). Product yields for the wildtype Tbb TIM and monoTIM-catalyzed reactions of glycolaldehyde labeled with carbon-13 at the carbonyl carbon ([1-13C]-GA) at pD 7.0 in the presence of phosphite dianion and in its absence were determined by 1H NMR spectroscopy [Go, M. K., Amyes, T. L., and Richard, J. P. (2009) Biochemistry 48, 5769–5778]. There is no detectable difference in the yields of the products of wildtype muscle and Tbb TIM-catalyzed reactions of [1-13C]-GA in D2O. The kinetic parameters for phosphite dianion activation of the reactions of [1-13C]-GA catalyzed by wildtype Tbb TIM are similar to those reported for the enzyme from rabbit muscle TIM [Amyes, T. L., and Richard, J. P. (2007), Biochemistry 46, 5841–5854], but there is no detectable dianion activation of the reaction catalyzed by monoTIM. The engineered disruption of subunit contacts at monoTIM causes movement of the essential side chains of Lys-13 and His-95 away from the catalytic active positions. We suggest that this places an increased demand that the intrinsic binding energy of phosphite dianion be utilized to drive the change in the conformation of monoTIM back to the active structure for wildtype TIM, with the result that there is insufficient binding energy remaining to give a detectable stabilization of the transition state for the monoTIM-catalyzed reaction of [1-13C]-GA.

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Carbohydrate metabolism

Opperdoes¹

1999

Progress in Human African Trypanosomiasis, Sleeping Sickness

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Selective Interaction of Glycosomal Enzymes from Trypanosoma brucei with Hydrophobic Cyclic Hexapeptides

Cited by 4 publications

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Synthetic peptides as inactivators of multimeric enzymes: inhibition of Plasmodium falciparum triosephosphate isomerase by interface peptides

Synthetic peptides as inactivators of multimeric enzymes: inhibition of Plasmodium falciparum triosephosphate isomerase by interface peptides

Wildtype and Engineered Monomeric Triosephosphate Isomerase from Trypanosoma brucei: Partitioning of Reaction Intermediates in D₂O and Activation by Phosphite Dianion

Carbohydrate metabolism

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Selective Interaction of Glycosomal Enzymes from Trypanosoma brucei with Hydrophobic Cyclic Hexapeptides

Cited by 4 publications

References 0 publications

Synthetic peptides as inactivators of multimeric enzymes: inhibition of Plasmodium falciparum triosephosphate isomerase by interface peptides

Synthetic peptides as inactivators of multimeric enzymes: inhibition of Plasmodium falciparum triosephosphate isomerase by interface peptides

Wildtype and Engineered Monomeric Triosephosphate Isomerase from Trypanosoma brucei: Partitioning of Reaction Intermediates in D2O and Activation by Phosphite Dianion

Carbohydrate metabolism

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Wildtype and Engineered Monomeric Triosephosphate Isomerase from Trypanosoma brucei: Partitioning of Reaction Intermediates in D₂O and Activation by Phosphite Dianion