In-gel activity-based protein profiling of a clickable covalent ERK1/2 inhibitor

被引：17

作者：

Lebraud, Honorine ^{[1
]}

Wright, David J. ^{[1
]}

East, Charlotte E. ^{[1
]}

Holding, Finn P. ^{[1
]}

O'Reilly, Marc ^{[1
]}

Heightman, Tom D. ^{[1
]}

机构：

[1] Astex Pharmaceut, 436 Cambridge Sci Pk, Cambridge CB4 0QA, England

来源：

MOLECULAR BIOSYSTEMS | 2016年 / 12卷 / 09期

关键词：

CHEMICAL PROBES; LIVING CELLS; LIVE CELLS; DISCOVERY; KINASE; TETRAZINE; SELECTIVITY; CHEMISTRY; MECHANISM; LIGATION;

D O I：

10.1039/c6mb00367b

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

In-gel activity-based protein profiling (ABPP) offers rapid assessment of the proteome-wide selectivity and target engagement of a chemical tool. Here we demonstrate the use of the inverse electron demand Diels Alder (IEDDA) click reaction for in-gel ABPP by evaluating the selectivity profile and target engagement of a covalent ERK1/2 probe tagged with a trans-cyclooctene group. The chemical probe was shown to bind covalently to Cys166 of ERK2 using protein MS and X-ray crystallography, and displayed submicromolar GI(50)s in A375 and HCT116 cells. In both cell lines, the probe demonstrated target engagement and a good selectivity profile at low concentrations, which was lost at higher concentrations. The IEDDA cycloaddition enabled fast and quantitative fluorescent tagging for readout with a high background-to-noise ratio and thereby provides a promising alternative to the commonly used copper catalysed alkyne-azide cycloaddition.

引用

页码：2867 / 2874

页数：8

共 33 条

[1] Structure-Guided Design of Potent and Selective Pyrimidylpyrrole Inhibitors of Extracellular Signal-Regulated Kinase (ERK) Using Conformational Control [J].

Aronov, Alex M. ;

Tang, Qing ;

Martinez-Botella, Gabriel ;

Bemis, Guy W. ;

Cao, Jingrong ;

Chen, Guanjing ;

Ewing, Nigel P. ;

Ford, Pamella J. ;

Germann, Ursula A. ;

Green, Jeremy ;

Hale, Michael R. ;

Jacobs, Marc ;

Janetka, James W. ;

Maltais, Francois ;

Markland, William ;

Namchuk, Mark N. ;

Nanthakumar, Suganthini ;

Poondru, Srinivasu ;

Straub, Judy ;

ter Haar, Ernst ;

Xie, Xiaoling .

JOURNAL OF MEDICINAL CHEMISTRY, 2009, 52 (20) :6362-6368

[2] The promise and peril of chemical probes [J].

Arrowsmith, Cheryl H. ;

Audia, James E. ;

Austin, Christopher ;

Baell, Jonathan ;

Bennett, Jonathan ;

Blagg, Julian ;

Bountra, Chas ;

Brennan, Paul E. ;

Brown, Peter J. ;

Bunnage, Mark E. ;

Buser-Doepner, Carolyn ;

Campbell, Robert M. ;

Carter, Adrian J. ;

Cohen, Philip ;

Copeland, Robert A. ;

Cravatt, Ben ;

Dahlin, Jayme L. ;

Dhanak, Dashyant ;

Edwards, Aled M. ;

Frye, Stephen V. ;

Gray, Nathanael ;

Grimshaw, Charles E. ;

Hepworth, David ;

Howe, Trevor ;

Huber, Kilian V. M. ;

Jin, Jian ;

Knapp, Stefan ;

Kotz, Joanne D. ;

Kruger, Ryan G. ;

Lowe, Derek ;

Mader, Mary M. ;

Marsden, Brian ;

Mueller-Fahrnow, Anke ;

Mueller, Susanne ;

O'Hagan, Ronan C. ;

Overington, John P. ;

Owen, Dafydd R. ;

Rosenberg, Saul H. ;

Roth, Brian ;

Ross, Ruth ;

Schapira, Matthieu ;

Schreiber, Stuart L. ;

Shoichet, Brian ;

Sundstrom, Michael ;

Superti-Furga, Giulio ;

Taunton, Jack ;

Toledo-Sherman, Leticia ;

Walpole, Chris ;

Walters, Michael A. ;

Willson, Timothy M. .

NATURE CHEMICAL BIOLOGY, 2015, 11 (08) :536-541

[3] Tetrahydropyrrolo-diazepenones as inhibitors of ERK2 kinase [J].

Bagdanoff, Jeffrey T. ;

Jain, Rama ;

Han, Wooseok ;

Zhu, Shejin ;

Madiera, Ann-Marie ;

Lee, Patrick S. ;

Ma, Xiaolei ;

Poon, Daniel .

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 2015, 25 (18) :3788-3792

[4] Tetrazine ligation: Fast bioconjugation based on inverse-electron-demand Diels-Alder reactivity [J].

Blackman, Melissa L. ;

Royzen, Maksim ;

Fox, Joseph M. .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2008, 130 (41) :13518-+

[5] Discovery of Novel, Dual Mechanism ERK Inhibitors by Affinity Selection Screening of an Inactive Kinase [J].

Deng, Yongqi ;

Shipps, Gerald W., Jr. ;

Cooper, Alan ;

English, Jessie M. ;

Annis, D. Allen ;

Carr, Donna ;

Nan, Yang ;

Wang, Tong ;

Zhu, Hugh Y. ;

Chuang, Cheng-Chi ;

Dayananth, Priya ;

Hruza, Alan W. ;

Xiao, Li ;

Jin, Weihong ;

Kirschmeier, Paul ;

Windsor, William T. ;

Samatar, Ahmed A. .

JOURNAL OF MEDICINAL CHEMISTRY, 2014, 57 (21) :8817-8826

[6] Development of a 18F-Labeled Tetrazine with Favorable Pharmacokinetics for Bioorthogonal PET Imaging [J].

Denk, Christoph ;

Svatunek, Dennis ;

Filip, Thomas ;

Wanek, Thomas ;

Lumpi, Daniel ;

Froehlich, Johannes ;

Kuntner, Claudia ;

Mikula, Hannes .

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2014, 53 (36) :9655-9659

[7] Tetrazine-Based Cycloadditions: Application to Pretargeted Live Cell Imaging [J].

Devaraj, Neal K. ;

Weissleder, Ralph ;

Hilderbrand, Scott A. .

BIOCONJUGATE CHEMISTRY, 2008, 19 (12) :2297-2299

[8] Tripping the switch fantastic: How a protein kinase cascade can convert graded inputs into switch-like outputs [J].

Ferrell, JE .

TRENDS IN BIOCHEMICAL SCIENCES, 1996, 21 (12) :460-466

[9] The Impact of Chemical Probes in Drug Discovery: A Pharmaceutical Industry Perspective [J].

Garbaccio, Robert M. ;

Parmee, Emma R. .

CELL CHEMICAL BIOLOGY, 2016, 23 (01) :10-17

[10]

Haq N., 2014, Patent No. [2014124230A2, 2014124230]

← 1 2 3 4 →