Thymoquinone and Poloxin are slow-irreversible inhibitors to human Polo-like kinase 1 Polo-box domain

Yin Zhou; Chen Jianhua; Peter H. Rehse

doi:10.1016/S1000-1948(10)60032-9

Thymoquinone and Poloxin are slow-irreversible inhibitors to human Polo-like kinase 1 Polo-box domain

Yin Zhou, Chen Jianhua, Peter H. Rehse

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

7 Scopus citations

Abstract

Objective: To provide a kinetic model(s) and reveal the mechanism of thymoquinone and Poloxin blocking an emerging anti-cancer target, human Polo-like kinase 1 (hPlk1) Polo-box domain (PBD). Methods: The binding kinetics was determined by using a fluorescence polarization based assay. The putative mechanism was examined with a competition test. Results: Thymoquinone follows a one-step binding with an association rate constant (k₁) of 6.635×10³ L·mol^-1·min^-1. Poloxin fit a two-step binding with a dissociation constant (Ki) of 118 μmol/L for the intermediate complex and its isomerization rate (k₄) of 0.131 5 min^-1 to form an irreversible adduct. No significant dissociation was observed for either ligand up to 13 h. The inhibitors responded insignificantly to the presence of Michael donors as hPlk1-PBD competitors. Conclusion: Thymoquinone and Poloxin are slow-tight ligands to the hPlk1-PBD with kinetic models distinct from each other. Michael addition as the mechanism is excluded.

Original language	English (US)
Pages (from-to)	136-142
Number of pages	7
Journal	Journal of Medical Colleges of PLA
Volume	25
Issue number	3
DOIs	https://doi.org/10.1016/S1000-1948(10)60032-9
State	Published - Jun 2010
Externally published	Yes

Keywords

Fluorescence polarization
Irreversible inhibitor
Kinetics
Polo-like kinase

ASJC Scopus subject areas

General Medicine

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10.1016/S1000-1948(10)60032-9

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title = "Thymoquinone and Poloxin are slow-irreversible inhibitors to human Polo-like kinase 1 Polo-box domain",

abstract = "Objective: To provide a kinetic model(s) and reveal the mechanism of thymoquinone and Poloxin blocking an emerging anti-cancer target, human Polo-like kinase 1 (hPlk1) Polo-box domain (PBD). Methods: The binding kinetics was determined by using a fluorescence polarization based assay. The putative mechanism was examined with a competition test. Results: Thymoquinone follows a one-step binding with an association rate constant (k1) of 6.635×103 L·mol-1·min-1. Poloxin fit a two-step binding with a dissociation constant (Ki) of 118 μmol/L for the intermediate complex and its isomerization rate (k4) of 0.131 5 min-1 to form an irreversible adduct. No significant dissociation was observed for either ligand up to 13 h. The inhibitors responded insignificantly to the presence of Michael donors as hPlk1-PBD competitors. Conclusion: Thymoquinone and Poloxin are slow-tight ligands to the hPlk1-PBD with kinetic models distinct from each other. Michael addition as the mechanism is excluded.",

keywords = "Fluorescence polarization, Irreversible inhibitor, Kinetics, Polo-like kinase",

author = "Yin Zhou and Chen Jianhua and Rehse, {Peter H.}",

note = "Funding Information: We thank Xue J. (Shanghai Medicilon Inc.) for assistance in medicinal and analytical chemistry and Gao H. for her help in molecular biology. We are grateful to the Shanghai Medicilon Inc. and Dr. Chen Chunlin for financial support. This work is a co-sponsored graduate research project by China Pharmaceutical University and Shanghai Medicilon Inc.",

year = "2010",

month = jun,

doi = "10.1016/S1000-1948(10)60032-9",

language = "English (US)",

volume = "25",

pages = "136--142",

journal = "Journal of Medical Colleges of PLA",

issn = "1000-1948",

publisher = "Elsevier Limited",

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TY - JOUR

T1 - Thymoquinone and Poloxin are slow-irreversible inhibitors to human Polo-like kinase 1 Polo-box domain

AU - Zhou, Yin

AU - Jianhua, Chen

AU - Rehse, Peter H.

N1 - Funding Information: We thank Xue J. (Shanghai Medicilon Inc.) for assistance in medicinal and analytical chemistry and Gao H. for her help in molecular biology. We are grateful to the Shanghai Medicilon Inc. and Dr. Chen Chunlin for financial support. This work is a co-sponsored graduate research project by China Pharmaceutical University and Shanghai Medicilon Inc.

PY - 2010/6

Y1 - 2010/6

N2 - Objective: To provide a kinetic model(s) and reveal the mechanism of thymoquinone and Poloxin blocking an emerging anti-cancer target, human Polo-like kinase 1 (hPlk1) Polo-box domain (PBD). Methods: The binding kinetics was determined by using a fluorescence polarization based assay. The putative mechanism was examined with a competition test. Results: Thymoquinone follows a one-step binding with an association rate constant (k1) of 6.635×103 L·mol-1·min-1. Poloxin fit a two-step binding with a dissociation constant (Ki) of 118 μmol/L for the intermediate complex and its isomerization rate (k4) of 0.131 5 min-1 to form an irreversible adduct. No significant dissociation was observed for either ligand up to 13 h. The inhibitors responded insignificantly to the presence of Michael donors as hPlk1-PBD competitors. Conclusion: Thymoquinone and Poloxin are slow-tight ligands to the hPlk1-PBD with kinetic models distinct from each other. Michael addition as the mechanism is excluded.

AB - Objective: To provide a kinetic model(s) and reveal the mechanism of thymoquinone and Poloxin blocking an emerging anti-cancer target, human Polo-like kinase 1 (hPlk1) Polo-box domain (PBD). Methods: The binding kinetics was determined by using a fluorescence polarization based assay. The putative mechanism was examined with a competition test. Results: Thymoquinone follows a one-step binding with an association rate constant (k1) of 6.635×103 L·mol-1·min-1. Poloxin fit a two-step binding with a dissociation constant (Ki) of 118 μmol/L for the intermediate complex and its isomerization rate (k4) of 0.131 5 min-1 to form an irreversible adduct. No significant dissociation was observed for either ligand up to 13 h. The inhibitors responded insignificantly to the presence of Michael donors as hPlk1-PBD competitors. Conclusion: Thymoquinone and Poloxin are slow-tight ligands to the hPlk1-PBD with kinetic models distinct from each other. Michael addition as the mechanism is excluded.

KW - Fluorescence polarization

KW - Irreversible inhibitor

KW - Kinetics

KW - Polo-like kinase

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Thymoquinone and Poloxin are slow-irreversible inhibitors to human Polo-like kinase 1 Polo-box domain

Abstract

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