Adenosine Kinase of T. b. Rhodesiense identified as the putative target of 4-[5-(4-phenoxyphenyl)-2H-pyrazol-3-yl]morpholine using chemical proteomics.
BACKGROUND: Human African trypanosomiasis (HAT), a major parasitic disease spread in Africa, urgently needs novel targets and new efficacious chemotherapeutic agents. Recently, we discovered that 4-[5-(4-phenoxyphenyl)-2H-pyrazol-3-yl]morpholine (compound 1) exhibits specific antitrypanosomal activi...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2009-01-01
|
| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | http://europepmc.org/articles/PMC2724708?pdf=render |
| _version_ | 1818150300839575552 |
|---|---|
| author | Sabine Kuettel Marc Mosimann Pascal Mäser Marcel Kaiser Reto Brun Leonardo Scapozza Remo Perozzo |
| author_facet | Sabine Kuettel Marc Mosimann Pascal Mäser Marcel Kaiser Reto Brun Leonardo Scapozza Remo Perozzo |
| author_sort | Sabine Kuettel |
| collection | DOAJ |
| description | BACKGROUND: Human African trypanosomiasis (HAT), a major parasitic disease spread in Africa, urgently needs novel targets and new efficacious chemotherapeutic agents. Recently, we discovered that 4-[5-(4-phenoxyphenyl)-2H-pyrazol-3-yl]morpholine (compound 1) exhibits specific antitrypanosomal activity with an IC(50) of 1.0 microM on Trypanosoma brucei rhodesiense (T. b. rhodesiense), the causative agent of the acute form of HAT. METHODOLOGY/PRINCIPAL FINDINGS: In this work we show adenosine kinase of T. b. rhodesiense (TbrAK), a key enzyme of the parasite purine salvage pathway which is vital for parasite survival, to be the putative intracellular target of compound 1 using a chemical proteomics approach. This finding was confirmed by RNA interference experiments showing that down-regulation of adenosine kinase counteracts compound 1 activity. Further chemical validation demonstrated that compound 1 interacts specifically and tightly with TbrAK with nanomolar affinity, and in vitro activity measurements showed that compound 1 is an enhancer of TbrAK activity. The subsequent kinetic analysis provided strong evidence that the observed hyperactivation of TbrAK is due to the abolishment of the intrinsic substrate-inhibition. CONCLUSIONS/SIGNIFICANCE: The results suggest that TbrAK is the putative target of this compound, and that hyperactivation of TbrAK may represent a novel therapeutic strategy for the development of trypanocides. |
| first_indexed | 2024-12-11T13:20:45Z |
| format | Article |
| id | doaj.art-a5c0844f212a43ac80244dc2f2c3f261 |
| institution | Directory Open Access Journal |
| issn | 1935-2727 1935-2735 |
| language | English |
| last_indexed | 2024-12-11T13:20:45Z |
| publishDate | 2009-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj.art-a5c0844f212a43ac80244dc2f2c3f2612022-12-22T01:05:50ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352009-01-0138e50610.1371/journal.pntd.0000506Adenosine Kinase of T. b. Rhodesiense identified as the putative target of 4-[5-(4-phenoxyphenyl)-2H-pyrazol-3-yl]morpholine using chemical proteomics.Sabine KuettelMarc MosimannPascal MäserMarcel KaiserReto BrunLeonardo ScapozzaRemo PerozzoBACKGROUND: Human African trypanosomiasis (HAT), a major parasitic disease spread in Africa, urgently needs novel targets and new efficacious chemotherapeutic agents. Recently, we discovered that 4-[5-(4-phenoxyphenyl)-2H-pyrazol-3-yl]morpholine (compound 1) exhibits specific antitrypanosomal activity with an IC(50) of 1.0 microM on Trypanosoma brucei rhodesiense (T. b. rhodesiense), the causative agent of the acute form of HAT. METHODOLOGY/PRINCIPAL FINDINGS: In this work we show adenosine kinase of T. b. rhodesiense (TbrAK), a key enzyme of the parasite purine salvage pathway which is vital for parasite survival, to be the putative intracellular target of compound 1 using a chemical proteomics approach. This finding was confirmed by RNA interference experiments showing that down-regulation of adenosine kinase counteracts compound 1 activity. Further chemical validation demonstrated that compound 1 interacts specifically and tightly with TbrAK with nanomolar affinity, and in vitro activity measurements showed that compound 1 is an enhancer of TbrAK activity. The subsequent kinetic analysis provided strong evidence that the observed hyperactivation of TbrAK is due to the abolishment of the intrinsic substrate-inhibition. CONCLUSIONS/SIGNIFICANCE: The results suggest that TbrAK is the putative target of this compound, and that hyperactivation of TbrAK may represent a novel therapeutic strategy for the development of trypanocides.http://europepmc.org/articles/PMC2724708?pdf=render |
| spellingShingle | Sabine Kuettel Marc Mosimann Pascal Mäser Marcel Kaiser Reto Brun Leonardo Scapozza Remo Perozzo Adenosine Kinase of T. b. Rhodesiense identified as the putative target of 4-[5-(4-phenoxyphenyl)-2H-pyrazol-3-yl]morpholine using chemical proteomics. PLoS Neglected Tropical Diseases |
| title | Adenosine Kinase of T. b. Rhodesiense identified as the putative target of 4-[5-(4-phenoxyphenyl)-2H-pyrazol-3-yl]morpholine using chemical proteomics. |
| title_full | Adenosine Kinase of T. b. Rhodesiense identified as the putative target of 4-[5-(4-phenoxyphenyl)-2H-pyrazol-3-yl]morpholine using chemical proteomics. |
| title_fullStr | Adenosine Kinase of T. b. Rhodesiense identified as the putative target of 4-[5-(4-phenoxyphenyl)-2H-pyrazol-3-yl]morpholine using chemical proteomics. |
| title_full_unstemmed | Adenosine Kinase of T. b. Rhodesiense identified as the putative target of 4-[5-(4-phenoxyphenyl)-2H-pyrazol-3-yl]morpholine using chemical proteomics. |
| title_short | Adenosine Kinase of T. b. Rhodesiense identified as the putative target of 4-[5-(4-phenoxyphenyl)-2H-pyrazol-3-yl]morpholine using chemical proteomics. |
| title_sort | adenosine kinase of t b rhodesiense identified as the putative target of 4 5 4 phenoxyphenyl 2h pyrazol 3 yl morpholine using chemical proteomics |
| url | http://europepmc.org/articles/PMC2724708?pdf=render |
| work_keys_str_mv | AT sabinekuettel adenosinekinaseoftbrhodesienseidentifiedastheputativetargetof454phenoxyphenyl2hpyrazol3ylmorpholineusingchemicalproteomics AT marcmosimann adenosinekinaseoftbrhodesienseidentifiedastheputativetargetof454phenoxyphenyl2hpyrazol3ylmorpholineusingchemicalproteomics AT pascalmaser adenosinekinaseoftbrhodesienseidentifiedastheputativetargetof454phenoxyphenyl2hpyrazol3ylmorpholineusingchemicalproteomics AT marcelkaiser adenosinekinaseoftbrhodesienseidentifiedastheputativetargetof454phenoxyphenyl2hpyrazol3ylmorpholineusingchemicalproteomics AT retobrun adenosinekinaseoftbrhodesienseidentifiedastheputativetargetof454phenoxyphenyl2hpyrazol3ylmorpholineusingchemicalproteomics AT leonardoscapozza adenosinekinaseoftbrhodesienseidentifiedastheputativetargetof454phenoxyphenyl2hpyrazol3ylmorpholineusingchemicalproteomics AT remoperozzo adenosinekinaseoftbrhodesienseidentifiedastheputativetargetof454phenoxyphenyl2hpyrazol3ylmorpholineusingchemicalproteomics |