title: Structural basis of reversine selectivity in inhibiting Mps1 more potently than aurora B kinase.
abstract: Monopolar spindle 1 (Mps1, also known as TTK) is a protein kinase crucial for ensuring that cell division progresses to anaphase only after all chromosomes are connected to spindle microtubules. Incomplete chromosomal attachment leads to abnormal chromosome counts in the daughter cells (aneuploidy), a condition common in many solid cancers. Therefore Mps1 is an established target in cancer therapy. Mps1 kinase inhibitors include reversine (2-(4-morpholinoanilino)-6-cyclohexylaminopurine), a promiscuous compound first recognized as an inhibitor of the Aurora B mitotic kinase. Here, we present the 3.0-Å resolution crystal structure of the Mps1 kinase domain bound to reversine. Structural comparison of reversine bound to Mps1 and Aurora B, indicates a similar binding pose for the purine moiety of reversine making three conserved hydrogen bonds to the protein main chain, explaining the observed promiscuity of this inhibitor. The cyclohexyl and morpholinoaniline moieties of reversine however, have more extensive contacts with the protein in Mps1 than in Aurora B. This is reflected both in structure-based docking energy calculations, and in new experimental data we present here, that both confirm that the affinity of reversine towards Mps1 is about two orders of magnitude higher than towards Aurora B. Thus, our data provides detailed structural understanding of the existing literature that argues reversine inhibits Mps1 more efficiently than Aurora B based on biochemical and in-cell assays. Proteins 2016; 84:1761-1766. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Division of Biochemistry, Netherlands Cancer Institute, 1066CX, Amsterdam, The Netherlands.
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Hiruma
authors.affiliation: Division of Cell Biology, Netherlands Cancer Institute, 1066CX, Amsterdam, The Netherlands.
authors.forename: Andre
authors.initials: A
authors.lastname: Koch
authors.affiliation: Division of Biochemistry, Netherlands Cancer Institute, 1066CX, Amsterdam, The Netherlands.
authors.forename: Shreya
authors.initials: S
authors.lastname: Dharadhar
authors.affiliation: Division of Biochemistry, Netherlands Cancer Institute, 1066CX, Amsterdam, The Netherlands.
authors.forename: Robbie P
authors.initials: RP
authors.lastname: Joosten
authors.affiliation: Division of Biochemistry, Netherlands Cancer Institute, 1066CX, Amsterdam, The Netherlands.
authors.forename: Anastassis
authors.identifier: 0000-0002-1151-6227
authors.initials: A
authors.lastname: Perrakis
mesh_terms: D000595:Amino Acid Sequence; D000970:Antineoplastic Agents / Q000737:chemistry*; D064107:Aurora Kinase B / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D008024:Ligands; D062105:Molecular Docking Simulation; D009025:Morpholines / Q000737:chemistry*; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D047428:Protein Kinase Inhibitors / Q000737:chemistry*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017433:Protein Structure, Secondary; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011687:Purines / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013329:Structure-Activity Relationship; D013816:Thermodynamics
chemical_list: D000970:Antineoplastic Agents; D018797:Cell Cycle Proteins; D008024:Ligands; D009025:Morpholines; D047428:Protein Kinase Inhibitors; D011687:Purines; D011994:Recombinant Proteins; D011505:Protein-Tyrosine Kinases; C576921:AURKB protein, human; D064107:Aurora Kinase B; D017346:Protein Serine-Threonine Kinases; C075840:TTK protein, human; C484369:2-(4-morpholinoanilino)-6-cyclohexylaminopurine
keywords: 2-(4-morpholinoanilino)-6-cyclohexylaminopurine; X-ray crystallography; microscale thermophoresis; mitotic kinase; reversine