Cooperativity between Orthosteric Inhibitors and Allosteric Inhibitor 8-Anilino-1-Naphthalene Sulfonic Acid (ANS) in Cyclin-Dependent Kinase 2

Erik B. Faber; Defeng Tian; David Burban; Nicholas M. Levinson; Jon E. Hawkinson; Gunda I. Georg

doi:10.1021/acschembio.0c00169

Cooperativity between Orthosteric Inhibitors and Allosteric Inhibitor 8-Anilino-1-Naphthalene Sulfonic Acid (ANS) in Cyclin-Dependent Kinase 2

Erik B. Faber, Defeng Tian, David Burban, Nicholas M. Levinson, Jon E. Hawkinson, Gunda I. Georg

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

While kinases have been attractive targets to combat many diseases, including cancer, selective kinase inhibition has been challenging, because of the high degree of structural homology in the active site, where many kinase inhibitors bind. We have previously discovered that 8-anilino-1-naphthalene sulfonic acid (ANS) binds an allosteric pocket in cyclin-dependent kinase 2 (Cdk2). Here, we detail the positive cooperativity between ANS and orthosteric Cdk2 inhibitors dinaciclib and roscovitine, which increase the affinity of ANS toward Cdk2 5-fold to 10-fold, and the relatively noncooperative effects of ATP. We observe these effects using a fluorescent binding assay and heteronuclear single quantum correlation nuclear magnetic resonance (HSQC NMR), where we noticed a shift from fast exchange to slow exchange upon ANS titration in the presence of roscovitine but not with an ATP mimic. The discovery of cooperative relationships between orthosteric and allosteric kinase inhibitors could further the development of selective kinase inhibitors in general.

Original language	English (US)
Pages (from-to)	1759-1764
Number of pages	6
Journal	ACS Chemical Biology
Volume	15
Issue number	7
DOIs	https://doi.org/10.1021/acschembio.0c00169
State	Published - Jul 17 2020

Bibliographical note

Funding Information:
We thank E. Scho?nbrunn for providing purified Cdk2. We thank S. Jakkaraj for providingp-Cl-ANS. This work was supported by NIH/NICHD 5 U01 HD080431, NIH/NICHD 1 R61 HD099743, and NIH/NIGMS R01 GM121515. E.F. was supported by NIH/NIGMS (through training Grant Nos. T32 GM008244 and T32 GM008700), as well as by a NIH/NCI fellowship (No. F30 CA232303).

Funding Information:
We thank E. Schönbrunn for providing purified Cdk2. We thank S. Jakkaraj for providing p-Cl-ANS. This work was supported by NIH/NICHD 5 U01 HD080431, NIH/NICHD 1 R61 HD099743, and NIH/NIGMS R01 GM121515. E.F. was supported by NIH/NIGMS (through training Grant Nos. T32 GM008244 and T32 GM008700), as well as by a NIH/NCI fellowship (No. F30 CA232303).

Publisher Copyright:
Copyright © 2020 American Chemical Society.

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Cooperativity between Orthosteric Inhibitors and Allosteric Inhibitor 8-Anilino-1-Naphthalene Sulfonic Acid (ANS) in Cyclin-Dependent Kinase 2. / Faber, Erik B.; Tian, Defeng ; Burban, David ; Levinson, Nicholas M.; Hawkinson, Jon E.; Georg, Gunda I.

In: ACS Chemical Biology, Vol. 15, No. 7, 17.07.2020, p. 1759-1764.

Research output: Contribution to journal › Article › peer-review

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title = "Cooperativity between Orthosteric Inhibitors and Allosteric Inhibitor 8-Anilino-1-Naphthalene Sulfonic Acid (ANS) in Cyclin-Dependent Kinase 2",

abstract = "While kinases have been attractive targets to combat many diseases, including cancer, selective kinase inhibition has been challenging, because of the high degree of structural homology in the active site, where many kinase inhibitors bind. We have previously discovered that 8-anilino-1-naphthalene sulfonic acid (ANS) binds an allosteric pocket in cyclin-dependent kinase 2 (Cdk2). Here, we detail the positive cooperativity between ANS and orthosteric Cdk2 inhibitors dinaciclib and roscovitine, which increase the affinity of ANS toward Cdk2 5-fold to 10-fold, and the relatively noncooperative effects of ATP. We observe these effects using a fluorescent binding assay and heteronuclear single quantum correlation nuclear magnetic resonance (HSQC NMR), where we noticed a shift from fast exchange to slow exchange upon ANS titration in the presence of roscovitine but not with an ATP mimic. The discovery of cooperative relationships between orthosteric and allosteric kinase inhibitors could further the development of selective kinase inhibitors in general.",

author = "Faber, {Erik B.} and Defeng Tian and David Burban and Levinson, {Nicholas M.} and Hawkinson, {Jon E.} and Georg, {Gunda I.}",

note = "Funding Information: We thank E. Scho?nbrunn for providing purified Cdk2. We thank S. Jakkaraj for providingp-Cl-ANS. This work was supported by NIH/NICHD 5 U01 HD080431, NIH/NICHD 1 R61 HD099743, and NIH/NIGMS R01 GM121515. E.F. was supported by NIH/NIGMS (through training Grant Nos. T32 GM008244 and T32 GM008700), as well as by a NIH/NCI fellowship (No. F30 CA232303). Funding Information: We thank E. Sch{\"o}nbrunn for providing purified Cdk2. We thank S. Jakkaraj for providing p-Cl-ANS. This work was supported by NIH/NICHD 5 U01 HD080431, NIH/NICHD 1 R61 HD099743, and NIH/NIGMS R01 GM121515. E.F. was supported by NIH/NIGMS (through training Grant Nos. T32 GM008244 and T32 GM008700), as well as by a NIH/NCI fellowship (No. F30 CA232303). Publisher Copyright: Copyright {\textcopyright} 2020 American Chemical Society.",

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AU - Hawkinson, Jon E.

AU - Georg, Gunda I.

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N2 - While kinases have been attractive targets to combat many diseases, including cancer, selective kinase inhibition has been challenging, because of the high degree of structural homology in the active site, where many kinase inhibitors bind. We have previously discovered that 8-anilino-1-naphthalene sulfonic acid (ANS) binds an allosteric pocket in cyclin-dependent kinase 2 (Cdk2). Here, we detail the positive cooperativity between ANS and orthosteric Cdk2 inhibitors dinaciclib and roscovitine, which increase the affinity of ANS toward Cdk2 5-fold to 10-fold, and the relatively noncooperative effects of ATP. We observe these effects using a fluorescent binding assay and heteronuclear single quantum correlation nuclear magnetic resonance (HSQC NMR), where we noticed a shift from fast exchange to slow exchange upon ANS titration in the presence of roscovitine but not with an ATP mimic. The discovery of cooperative relationships between orthosteric and allosteric kinase inhibitors could further the development of selective kinase inhibitors in general.

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Cooperativity between Orthosteric Inhibitors and Allosteric Inhibitor 8-Anilino-1-Naphthalene Sulfonic Acid (ANS) in Cyclin-Dependent Kinase 2

Abstract

Bibliographical note

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