Intracellular targets of cyclin-dependent kinase inhibitors: identification by affinity chromatography using immobilised inhibitors.

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Intracellular targets of cyclin-dependent kinase inhibitors: identification by affinity chromatography using immobilised inhibitors. / Knockaert, Marie; Gray, N.; Damiens, E. et al.
In: Chemistry and Biology, Vol. 7, No. 6, 01.06.2000, p. 411-422.

Research output: Contribution to Journal/Magazine › Journal article › peer-review

Harvard

Knockaert, M, Gray, N, Damiens, E, Chang, Y-T, Grellier, P, Grant, KM, Fergusson, D, Mottram, JC, Soete, M, Dubremetz, J-F, Le Roch, K, Doerig, C, Schultz, PG & Meijer, L 2000, 'Intracellular targets of cyclin-dependent kinase inhibitors: identification by affinity chromatography using immobilised inhibitors.', Chemistry and Biology, vol. 7, no. 6, pp. 411-422. https://doi.org/10.1016/S1074-5521(00)00124-1

APA

Knockaert, M., Gray, N., Damiens, E., Chang, Y.-T., Grellier, P., Grant, K. M., Fergusson, D., Mottram, J. C., Soete, M., Dubremetz, J.-F., Le Roch, K., Doerig, C., Schultz, P. G., & Meijer, L. (2000). Intracellular targets of cyclin-dependent kinase inhibitors: identification by affinity chromatography using immobilised inhibitors. Chemistry and Biology, 7(6), 411-422. https://doi.org/10.1016/S1074-5521(00)00124-1

Vancouver

Knockaert M, Gray N, Damiens E, Chang YT, Grellier P, Grant KM et al. Intracellular targets of cyclin-dependent kinase inhibitors: identification by affinity chromatography using immobilised inhibitors. Chemistry and Biology. 2000 Jun 1;7(6):411-422. doi: 10.1016/S1074-5521(00)00124-1

Author

Knockaert, Marie ; Gray, N. ; Damiens, E. et al. / Intracellular targets of cyclin-dependent kinase inhibitors: identification by affinity chromatography using immobilised inhibitors. In: Chemistry and Biology. 2000 ; Vol. 7, No. 6. pp. 411-422.

Bibtex

@article{74cb0803a0d14f9c89d5f0443fd59b25,

title = "Intracellular targets of cyclin-dependent kinase inhibitors: identification by affinity chromatography using immobilised inhibitors.",

abstract = "Background: Chemical inhibitors of cyclin-dependent kinases (CDKs) have great therapeutic potential against various proliferative and neurodegenerative disorders. Olomoucine, a 2,6,9-trisubstituted purine, has been optimized for activity against CDK1/cyclin B by combinatorial and medicinal chemistry efforts to yield the purvalanol inhibitors. Although many studies support the action of purvalanols against CDKs, the actual intracellular targets of 2,6,9-trisubstituted purines remain unverified. Results: To address this issue, purvalanol B (95) and an N6-methylated, CDK-inactive derivative (95M) were immobilized on an agarose matrix. Extracts from a diverse collection of cell types and organisms were screened for proteins binding purvalanol B. In addition to validating CDKs as intracellular targets, a variety of unexpected protein kinases were recovered from the 95 matrix. Casein kinase 1 (CK1) was identified as a principal 95 matrix binding protein in Plasmodium falciparum, Leishmania mexicana, Toxoplasma gondii and Trypanosoma cruzi. Purvalanol compounds also inhibit the proliferation of these parasites, suggesting that CK1 is a valuable target for further screening with 2,6,9-trisubstituted purine libraries. Conclusions: That a simple batchwise affinity chromatography approach using two purine derivatives facilitated isolation of a small set of highly purified kinases suggests that this could be a general method for identifying intracellular targets relevant to a particular class of ligands. This method allows a close correlation to be established between the pattern of proteins bound to a small family of related compounds and the pattern of cellular responses to these compounds.",

keywords = "Casein kinase 1, Cyclin-dependent kinases, Erk, Malaria, Purine",

author = "Marie Knockaert and N. Gray and E. Damiens and Y.-T. Chang and P. Grellier and Grant, {Karen M.} and David Fergusson and Mottram, {Jeremy C.} and M. Soete and J.-F. Dubremetz and {Le Roch}, K. and C. Doerig and Schultz, {P. G.} and Laurent Meijer",

year = "2000",

month = jun,

day = "1",

doi = "10.1016/S1074-5521(00)00124-1",

language = "English",

volume = "7",

pages = "411--422",

journal = "Chemistry and Biology",

issn = "1074-5521",

publisher = "Cell Press",

number = "6",

}

RIS

TY - JOUR

T1 - Intracellular targets of cyclin-dependent kinase inhibitors: identification by affinity chromatography using immobilised inhibitors.

AU - Knockaert, Marie

AU - Gray, N.

AU - Damiens, E.

AU - Chang, Y.-T.

AU - Grellier, P.

AU - Grant, Karen M.

AU - Fergusson, David

AU - Mottram, Jeremy C.

AU - Soete, M.

AU - Dubremetz, J.-F.

AU - Le Roch, K.

AU - Doerig, C.

AU - Schultz, P. G.

AU - Meijer, Laurent

PY - 2000/6/1

Y1 - 2000/6/1

N2 - Background: Chemical inhibitors of cyclin-dependent kinases (CDKs) have great therapeutic potential against various proliferative and neurodegenerative disorders. Olomoucine, a 2,6,9-trisubstituted purine, has been optimized for activity against CDK1/cyclin B by combinatorial and medicinal chemistry efforts to yield the purvalanol inhibitors. Although many studies support the action of purvalanols against CDKs, the actual intracellular targets of 2,6,9-trisubstituted purines remain unverified. Results: To address this issue, purvalanol B (95) and an N6-methylated, CDK-inactive derivative (95M) were immobilized on an agarose matrix. Extracts from a diverse collection of cell types and organisms were screened for proteins binding purvalanol B. In addition to validating CDKs as intracellular targets, a variety of unexpected protein kinases were recovered from the 95 matrix. Casein kinase 1 (CK1) was identified as a principal 95 matrix binding protein in Plasmodium falciparum, Leishmania mexicana, Toxoplasma gondii and Trypanosoma cruzi. Purvalanol compounds also inhibit the proliferation of these parasites, suggesting that CK1 is a valuable target for further screening with 2,6,9-trisubstituted purine libraries. Conclusions: That a simple batchwise affinity chromatography approach using two purine derivatives facilitated isolation of a small set of highly purified kinases suggests that this could be a general method for identifying intracellular targets relevant to a particular class of ligands. This method allows a close correlation to be established between the pattern of proteins bound to a small family of related compounds and the pattern of cellular responses to these compounds.

AB - Background: Chemical inhibitors of cyclin-dependent kinases (CDKs) have great therapeutic potential against various proliferative and neurodegenerative disorders. Olomoucine, a 2,6,9-trisubstituted purine, has been optimized for activity against CDK1/cyclin B by combinatorial and medicinal chemistry efforts to yield the purvalanol inhibitors. Although many studies support the action of purvalanols against CDKs, the actual intracellular targets of 2,6,9-trisubstituted purines remain unverified. Results: To address this issue, purvalanol B (95) and an N6-methylated, CDK-inactive derivative (95M) were immobilized on an agarose matrix. Extracts from a diverse collection of cell types and organisms were screened for proteins binding purvalanol B. In addition to validating CDKs as intracellular targets, a variety of unexpected protein kinases were recovered from the 95 matrix. Casein kinase 1 (CK1) was identified as a principal 95 matrix binding protein in Plasmodium falciparum, Leishmania mexicana, Toxoplasma gondii and Trypanosoma cruzi. Purvalanol compounds also inhibit the proliferation of these parasites, suggesting that CK1 is a valuable target for further screening with 2,6,9-trisubstituted purine libraries. Conclusions: That a simple batchwise affinity chromatography approach using two purine derivatives facilitated isolation of a small set of highly purified kinases suggests that this could be a general method for identifying intracellular targets relevant to a particular class of ligands. This method allows a close correlation to be established between the pattern of proteins bound to a small family of related compounds and the pattern of cellular responses to these compounds.

KW - Casein kinase 1

KW - Cyclin-dependent kinases

KW - Erk

KW - Malaria

KW - Purine

U2 - 10.1016/S1074-5521(00)00124-1

DO - 10.1016/S1074-5521(00)00124-1

M3 - Journal article

VL - 7

SP - 411

EP - 422

JO - Chemistry and Biology

JF - Chemistry and Biology

SN - 1074-5521

IS - 6

ER -

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