Technology Overview

Expert Opin Ther Targets. 2006;10(1):179-190.
Transcreener™: screening enzymes involved in covalent regulation
Robert G Lowery & Karen Kleman-Leyer

Expert opinion on drug discovery. 2011 Jan 1;6(1):17-32.
Flurorescence polarization assays in small molecule screening.
Lea WA, Simeonov A.

ADP Assay

bioRxiv. 2022 July 30.
Molecular basis for differential activation of p101 and p84 complexes of PI3Kγ by Ras and GPCRs.  
Rathinaswamy M, Jenkins M, Zhang X, Stariha J, Ranga-Prasad H, Dalwadi U, Fleming K, Yip C, Williams R, Burke J.

bioRxiv. 2022 July 20.
Epinephrine inhibits PI3K alpha via the Hippo kinases. 
Lin T, Ramsamooj S, Liberatore K, Lantier L, Vasan N, Karukurichi K, Hwang S, Kesicki E, Kastenhuber E, Weiderhold T, Yaron T, Zhu M, Ma Y, Paddock M, Zhang G, Hopkins B, McGuinness O, Schwartz R, Cantley L, Johnson J, Goncalves M.

bioRxiv. 2022 April 6.
Oncogenic mutations of PIK3CA lead to increased membrane of recruitment driven by reorientation of the ABD, p85, and C-terminus. 
Ranga-Prasad H, Jenkins M, Parson M, Rathinaswamy M, Burke J.

Nature Communications. 2022 Jan 28; 13(569).
Decisive role of water and protein dynamics in residence time of p38α MAP kinase inhibitors. 
Pantsar T, Kaiser P, Kudolo M, Forster M, Rothbauer U, Laufer S.

Science Advances. 2021 Aug 27; 7(35).
Structure of the phosphoinositide 3-kinase (PI3K) p110y-p101 complex reveals molecular mechanism of GPCR activation. 
Rathinaswamy M, Dalwadi U, Fleming K, Adams C, Stariha J, Pardon E, Baek M, Vadas O, Dimaio F, Burke J.

Biomolecules. 2021 Aug 12; 11(8):1196.
Structural and Functional Insights into the Biofilm-Associated BceF Tyrosine Kinase Domain Burkholderia cepacia. 
Mayer M, Matiuhin Y, Nawatha M, Tabachnikov O, Fish I, Schultz N, Dvir H, Landau M.

Science Advances. 2021 Aug 6; 7(32).
The structural basis of bacterial manganese import. 
Neville S, JÖHAMN J, Watts J, Macdermott-Opeskin H, Fairweather S, Ganio K, Hulyer A, McGrath A, Hayes A, Malcom T, Davies M, Nomura N, Iwata S, O'mara M, Maher M, McDevitt C.

ChemBioChem. 2021 July 22; 22:3236-3253.
Enzymatic Assays to Explore Viral mRNA Capping Machinery. 
Kasprzyk R, Jemielity J.

ChemRxiv. 2021 July 20.
Jaeger-Hunting for Anti-Malarials with Generative Chemistry.
Godinez W, Ma E, Chao A, Pei L, Skewes-Cox P, Canham S, Jenkins J, Young J, Martin E, Guiguemde A.

eLife. 2021 Mar 4; 10:e64691.
Disease-related mutations in PI3Ky disrupt regulatory C-terminal dynamics and reveal a path to selective inhibitors.
Rathinaswamy M, Gaieb Z, Fleming K, Borsari C, Harris N, Moeller B, Wymann M, Amaro R, Burke J.

eLife. 2021 Feb 19; 10:e63524.
Molecular Structures of the Eukaryotic Retinal Importer ABCA4.
Liu F, Lee J, Chen J.

Cancer Research. 2020 Dec 2; 80(23):5270-5281.
A Novel Inhibitor of HSP70 Induces Mitochondrial Toxicity and Immune Cell Recruitment in Tumors.
Barnoud T, Leung J, I-Ju Leu J, Basu S, Narayana Reddy Poli A, Parris J, Indeglia A, Martynyuk T, Good M, Gnanapradeepan A, Liu Q, Speicher D, Gabrilovich D, Salvino J, George D, Murphy M.

Nature Communications. 2020 Oct 20; 11(5237)
CHIP phosphorylation by protein kinase G enhances protein quality control and attenuates cardiac ischemic injury.
Ranek M, Oeing C, Sanchez-Hodge R, Kokkonen-Simon K, Dillard D, Aslam I, Rainer P, Mishra S, Dunkerly-Erying B, Holewinski R, Virus C, Zhang H, Mannion M, Agrawal V, Hahn V, Lee D, Sasaki M, Van Eyk K, Willis M, Page R, Schisler J, Kass D.

Cell Chemical Biology. 2020 Feb 3.
Small Molecule Inhibition of CPS1 Activity through an Allosteric Pocket.
Yao S, Nguyen TV, Rolfe A, Agrawal AA, Ke J, Peng S, Colombo F, Yu S, Bouchard P, Wu J, Huang KC.

Science. 2019 Nov 8;366(6466):714-23.
Double PIK3CA mutations in cis increase oncogenicity and sensitivity to PI3Kα inhibitors. 
Vasan N, Razavi P, Johnson JL, Shao H, Shah H, Antoine A, Ladewig E, Gorelick A, Lin TY, Toska E, Xu G.

Bioorganic & medicinal chemistry letters. 2019 Jun 15;29(12):1497-501.
Potent and selective inhibitors of receptor-interacting protein kinase 1 that lack an aromatic back pocket group.
Hamilton GL, Chen H, Deshmukh G, Eigenbrot C, Fong R, Johnson A, Kohli PB, Lupardus PJ, Liederer BM, Ramaswamy S, Wang H.

bioRxiv. 2019 Jan 1:634592.
Characterisation of the c10orf76-PI4KB complex, and its necessity for Golgi PI4P levels and enterovirus replication. 
McPhail JA, Lyoo H, Pemberton JG, Hoffmann RM, van Elst W, Strating JR, Jenkins ML, Stariha J, van Kuppeveld FJ, Balla T, Burke JE.

Nature. 2018 May;557(7704):196.
Structural basis for dual-mode inhibition of the ABC transporter MsbA.
Ho H, Miu A, Alexander MK, Garcia NK, Oh A, Zilberleyb I, Reichelt M, Austin CD, Tam C, Shriver S, Hu H.

Antimicrobial agents and chemotherapy. 2018 Apr 1;62(4):e02151-17.
A novel inhibitor of the LolCDE ABC transporter essential for lipoprotein trafficking in Gram-negative bacteria. 
Nickerson NN, Jao CC, Xu Y, Quinn J, Skippington E, Alexander MK, Miu A, Skelton N, Hankins JV, Lopez MS, Koth CM.

SLAS DISCOVERY: Advancing Life Sciences R&D. 2018 Jan;23(1):11-22.
A High-throughput screening triage workflow to authenticate a novel series of PFKFB3 inhibitors.
St-Gallay SA, Bennett N, Critchlow SE, Curtis N, Davies G, Debreczeni J, Evans N, Hardern I, Holdgate G, Jones NP, Leach L.

SLAS DISCOVERY: Advancing Life Sciences R&D. 2017 Feb 1:2472555217695080.
A High-Throughput Method for Measuring Drug Residence Time Using the Transcreener ADP Assay.
Kumar M, Lowery RG.

Journal of Biomolecular Screening. 2016 Oct;21(9):912-22.
Development of a 3D Tissue Culture–Based High-Content Screening Platform That Uses Phenotypic Profiling to Discriminate Selective Inhibitors of Receptor Tyrosine Kinases.
Booij TH, Klop MJ, Yan K, Szántai-Kis C, Szokol B, Orfi L, van de Water B, Keri G, Price LS.

PloS one. 2015 Apr 14;10(4):e0124234.
Targeting Vascular Endothelial Growth Factor Receptor 2 and Protein Kinase D1 Related Pathways by a Multiple Kinase Inhibitor in Angiogenesis and Inflammation Related Processes In Vitro.
Varga A, Gyulavári P, Greff Z, Futosi K, Németh T, Simon-Szabó L, Kerekes K, Szántai-Kis C, Brauswetter D, Kokas M, Borbély G.

Analytical Methods. 2014;6(5):1468-74.
Development of a sensitive assay for SERCA activity using FRET detection of ADP.
Jing M, Verardi R, Veglia G, Bowser MT.

Molecular cancer therapeutics. 2014 Jun 1;13(6):1442-56.
A novel CDK9 inhibitor shows potent antitumor efficacy in preclinical hematologic tumor models.
Yin T, Lallena MJ, Kreklau EL, Fales KR, Carballares S, Torrres R, Wishart GN, Ajamie RT, Cronier DM, Iversen PW, Meier TI.

Biochemical Journal. 2013 Aug 15;454(1):49-57.
Evolutionarily conserved structural changes in phosphatidylinositol 5-phosphate 4-kinase (PI5P4K) isoforms are responsible for differences in enzyme activity and localization.
Clarke JH, Irvine RF.

J Biol Chem. 2013 May 23. [Epub ahead of print]
Primuline Derivatives That Mimic RNA To Stimulate Hepatitis C Virus NS3 Helicase-Catalyzed ATP Hydrolysis.
Sweeney NL, Shadrick WR, Mukherjee S, Li K, Frankowski KJ, Schoenen FJ, Frick DN.

J Biomol Screen. 2013 Mar 27. [Epub ahead of print]
Discovering New Medicines Targeting Helicases: Challenges and Recent Progress.
Shadrick WR, Ndjomou J, Kolli R, Mukherjee S, Hanson AM, Frick DN.

Advances in Biological Regulation. 2013 Jan 31;53(1):97-110.
Dynamic steps in receptor tyrosine kinase mediated activation of class IA phosphoinositide 3-kinases (PI3K) captured by H/D exchange (HDX-MS).
Burke JE, Williams RL.

Assay Drug Dev Technol. 2012 Jun;10(3):260-8. Epub 2011 Dec 22.
High-Throughput Screening for RecA Inhibitors Using a Transcreener Adenosine 5′-O-Diphosphate Assay.
Peterson EJ, Janzen WP, Kireev D, Singleton SF.

Mol Cell. 2011 Mar 4;41(5):567-78.
Structure of lipid kinase p110ß/p85ß elucidates an unusual SH2-domain-mediated inhibitory mechanism.
Zhang X, Vadas O, Perisic O, Anderson KE, Clark J, Hawkins PT, Stephens LR, Williams RL.

Mol Biochem Parasitol. 2011 Jan: 175(1):21-29
Identification of inhibitors for putative malaria drug targets among novel antimalarial compounds
Crowther GJ, Napuli AJ, Gilligan JH, Gagaring K, Borboa R, Francek C, Chen Z, Dagostino EF, Stockmyer JB, Wang Y, Rodenbough PP, Castaneda LJ, Leibly DJ, Bhandari J, Gelb MH, Brinker A, Engels IH, Taylor J, Chatterjee AK, Fantauzzi P, Glynne RJ, Van Voorhis WC, Kuhen KL.

J Biomol Screen. 2010 Mar;15(3):279-86. Epub 2010 Feb 10.
Detection of the ATPase activity of the molecular chaperones Hsp90 and Hsp72 using the Transcreener ADP assay kit.
Rowlands M, McAndrew C, Prodromou C, Pearl L, Kalusa A, Jones K, Workman P, Aherne W.

J Biomol Screen. 2009; 14(7): 838-844
High-Throughput, Cell-Free, Liposome-Based Approach for Assessing In Vitro Activity of Lipid Kinases
Demian DJ, Clugston SL, Foster MM, Rameh L, Sarkes D, Townson SA, Yang L, Zhang M, Charlton ME

J Biomol Screen. 2009; 14(6): 679-689
Efficient Elimination of Nonstoichiometric Enzyme Inhibitors from HTS Hit Lists
Habig M, Blechschmidt A, Dressler S, Hess B, Patel V, Billich A, Ostermeier C, Beer D, Klumpp M.

Comb Chem High Throughput Screen 2009; 12(3):258-68
Development and Validation of a High-density Fluorescence Polarization-based Assay for the Trypanosoma RNA Triphosphatase TbCet1
Antczak C, Shum D, Radu C, Seshan VE, Djaballah H.

Assay Drug Dev Technol. 2009; 7(1):56-67
Characterization and Optimization of a Red-Shifted Fluorescence Polarization ADP Detection Assay
Kleman-Leyer KM, Klink TA, Kopp AL, Westermeyer TA, Koeff MD, Larson BR, Worzella TJ, Pinchard CA, van de Kar SAT, Zaman GJR, Hornberg JJ, Lowery RG.

J Biomol Screen. 2008; 13(6):476-485
Evaluating PI3 Kinase Isoforms Using Transcreener™ ADP Assays
Klink TA, Kleman-Leyer K, Kopp A, Westermeyer TA, Lowery RG.

J Biomol Screen. 2007; 12(4):578-584
Development of a Transcreener™ Kinase Assay for Protein Kinase A and Demonstration of concordance of Data with a Filter-Binding Assay Format
Huss KL, Blonigen PE, Campbell RM.

Assay Drug Dev Technol. 2007; 5:225-235
Discovery of Acetyl-Coenzyme A Carboxylase 2 Inhibitors: Comparison of a Fluorescence Intensity-Based Phosphate Assay and a Fluorescence Polarization-Based ADP Assay for High-Throughput Screening
Liu Y, Zalameda L, Kim KW, Wang M, McCarter JD.

AMP/GMP Assay

ACS Omega. 2021 Aug 12; 6(33):21,741-21,754.
On the Viability of Tadalafil-Based ¹⁸F-Radiotracers for In Vivo Phosphodiesterase 5 (PDE5) PET Imaging.
Bailey J, Wuest M, Bojovic T, Kronemann T, Wängler C, Wängler B. Wuest F, Schirrmacher R.

Cancer Res. 2021 Mar 1; 81(5);1252-1264.
Targeting ACSS2 with a Transition-State Mimetic Inhibits Triple-Negative Breast Cancer Growth.
Miller K, Pniewski K, Perry C, Papp S, Shaffer J, Velasco-Silva J, Casciano J, Aramburu T, Cassel S, Skordalakes E, Kossenkov A, Salvino J, Schug Z.

SLAS Discovery: Advancing the Science of Drug Discovery. 2021 Jan 5 :2472555220982321.
Development of a High-Throughput Assay to Identify Inhibitors of ENPP1.
Kumar M, Lowery RG.

Biomolecules. 2020 Dec 1; 10(12):1625.
Recent Development of Aminoacyl-tRNA Synthetase Inhibitors for Human Disease: A Future Perspective.
Kim S-H, Bae S, Song M.

SLAS DISCOVERY: Advancing Life Sciences R&D. 2019 Dec 22:2472555219893632.
High-Throughput Screening Assays for Cancer Immunotherapy Targets: Ectonucleotidases CD39 and CD73. 
Kumar M, Lowery R, Kumar V.

Analyst. 2015;140(1):140-8.
Fluorescence polarization immunoassays for monitoring nucleoside triphosphate diphosphohydrolase (NTPDase) activity.
Fiene A, Baqi Y, Lecka J, Sévigny J, Müller CE.

Biochemical Pharmacology. 2015 Jan 15;93(2):171-81.
Polyoxometalates—potent and selective ecto-nucleotidase inhibitors.
Lee SY, Fiene A, Li W, Hanck T, Brylev KA, Fedorov VE, Lecka J, Haider A, Pietzsch HJ, Zimmermann H, Sévigny J.

Journal of Biomolecular Screening. 2015 Jan;20(1):122-30.
Identification of potent inhibitors of the Trypanosoma brucei methionyl-tRNA synthetase via high-throughput orthogonal screening.
Pedró-Rosa L, Buckner FS, Ranade RM, Eberhart C, Madoux F, Gillespie JR, Koh CY, Brown S, Lohse J, Verlinde CL, Fan E.

Cell Host Microbe. 2012 Jun 14;11(6):654-63. doi: 10.1016/j.chom.2012.04.015.
Selective and specific inhibition of the plasmodium falciparum lysyl-tRNA synthetase by the fungal secondary metabolite cladosporin.
Hoepfner D, McNamara CW, Lim CS, Studer C, Riedl R, Aust T, McCormack SL, Plouffe DM, Meister S, Schuierer S, Plikat U, Hartmann N, Staedtler F, Cotesta S, Schmitt EK, Petersen F, Supek F, Glynne RJ, Tallarico JA, Porter JA, Fishman MC, Bodenreider C, Diagana TT, Movva NR, Winzeler EA.

J Biomol Screen. January 2012 vol. 17 no. 1 59-70
Development and Validation of a Generic Fluorescent Methyltransferase Activity Assay Based on the Transcreener AMP/GMP Assay.
Klink TA, Staeben M, Twesten K, Kopp AL, Kumar M, Schall Dunn R, Pinchard CA, Kleman-Leyer KM, Klumpp M, Lowery RG.

Science. 2011 Jul 22;333(6041):453-6
De-AMPylation of the small GTPase Rab1 by the pathogen Legionella pneumophila.
Neunuebel MR, Chen Y, Gaspar AH, Backlund PS Jr, Yergey A, Machner MP.

Assay Drug Dev Technol. 2010 Jun;8(3):344-55
Development and Validation of a Transcreener Assay for Detection of AMP- and GMP-Producing Enzymes
Staeben M, Kleman-Leyer KM, Kopp AL, Westermeyer TA, Lowery RG.

GDP Assay

Journal of Experimental Botany. 2020 Dec 30; 71(22):6932-6944.
Dissecting the role of a plant-specific Rab5 small GTPase NbRabF1 in Bamboo moasic virus infection.
Huang Y, Hou P, Chen I, Hsu Y, Tsai C, Cheng C.

The Journal of Infectious Diseases. 2020 Oct 1; 222(7):1188-1198.
The Stringent Response Contributes to Persistent Methicillin-Resistant Staphylococcus aureus Endovascular Infection Through the Purine Biosynthetic Pathway.
Li L, Bayer A, Cheung A, Lu L, Abdelhady W, Donegan N, Hong J, Yeaman M, Xiong Y.

Cells. 2020 April 22; 9(4):1042.
Molecular Regulation of the RhoGAP GRAF3 and Its Capacity to Limit Blood Pressure In Vivo.
Dee R, Bai X, Mack C, Taylor J.

Journal of Biological Chemistry. 2019 Nov 8;294(45):16865-83.
L-type prostaglandin D synthase regulates the trafficking of the PGD2 DP1 receptor by interacting with the GTPase Rab4.
Binda C, Génier S, Degrandmaison J, Picard S, Fréchette L, Jean S, Marsault E, Parent JL.

Methods in Molecular Biology, 2018. vol 1821. Humana Press, New York, NY
High-Throughput Assay for RhoGEFs Based on the Transcreener^® GDP Assay. In: Rivero F. (eds) Rho GTPases.
Lowery R.G., Kumar M.

PloS one. 2017 Sep 28;12(9):e0185639.
A highly-sensitive high throughput assay for dynamin’s basal GTPase activity.
Mohanakrishnan A, Tran TV, Kumar M, Chen H, Posner BA, Schmid SL.

Journal of Virology. 2015 Nov 1;89(21):10879-90.
Functional comparison of Mx1 from two different mouse species reveals the involvement of loop L4 in the antiviral activity against influenza A viruses.
Verhelst J, Spitaels J, Nürnberger C, De Vlieger D, Ysenbaert T, Staeheli P, Fiers W, Saelens X.

J Biomol Screen. 2015 Jul 20. pii: 1087057115596326. [Epub ahead of print]
A High-Throughput Assay for Rho Guanine Nucleotide Exchange Factors Based on the Transcreener GDP Assay
Reichman M, Schabdach A, Kumar M, Zielinski T, Donover PS, Laury-Kleintop LD, Lowery RG.

Curr Protoc Pharmacol. 2012 Mar;Chapter 2:Unit2.8.
Evaluating Modulators of “Regulator of G-protein Signaling” (RGS) Proteins.
Bosch DE, Zielinski T, Lowery RG, Siderovski DP.

J Biomol Screen. 2011 Aug;16(7):717-23. Epub 2011 May 18
High-throughput fluorescence polarization assay for the enzymatic activity of GTPase-activating protein of ADP-ribosylation factor (ARFGAP).
Sun W, Vanhooke JL, Sondek J, Zhang Q.

J Biomol Screen. 2009.
Two Gα_i1 Rate-Modifying Mutations Act in Concert to Allow Receptor-Independent, Steady-State Measurements of RGS Protein Activity
Zielinski T, Kimple AJ, Hutsell SQ, Koeff MD, Siderovski DP, Lowery RG.

EPIGEN Methyltransferase Assay

Assay and Drug Development Technologies. Jun 2015. 13(4), pp.200-209.
Biochemical Assay Development for Histone Methyltransferases Using a Transcreener-Based Assay for S-Adenosylhomocysteine
Kumar M, Zielinski T, Lowery RG.

UDP Assay

Antimicrobial agents and chemotherapy. 2018 May 1;62(5):e00107-18.
Treatment of Clostridium difficile infection with a small-molecule inhibitor of toxin UDP-glucose hydrolysis activity.
Stroke IL, Letourneau JJ, Miller TE, Xu Y, Pechik I, Savoly DR, Ma L, Sturzenbecker LJ, Sabalski J, Stein PD, Webb ML.

Assay and Drug Development Technologies. 2016. 14, no. 4 : 240-251.
Development and Validation of a Universal High-Throughput UDP-Glycosyltransferase Assay with a Time-Resolved FRET Signal.
Zielinski T, Reichman M, Donover PS, Lowery RG.

The Journal of Immunology. 2016 Feb 15;196(4):1790-8.
TLR-Activated Gap Junction Channels Protect Mice against Bacterial Infection through Extracellular UDP Release.
Qin J, Zhang G, Zhang X, Tan B, Lv Z, Liu M, Ren H, Qian M, Du B.

The Journal of Immunology. 2014 Nov 1;193(9):4515-26.
Extracellular UDP and P2Y6 function as a danger signal to protect mice from vesicular stomatitis virus infection through an increase in IFN-β production.
Li R, Tan B, Yan Y, Ma X, Zhang N, Zhang Z, Liu M, Qian M, Du B.

Chembiochem. 2010 Sep 24;11(14):1939-49. Review
Glycosyltransferases and their assays.
Wagner GK, Pesnot T.

cGAMP Assay

Bone Research. Feb 24; 10(19).
RIOX1-Demethylated cGAS Regulates Ionizing Radiation-Elicited DNA Repair.
Xiao Y, Li J, Liao X, He Y, He T, Yang C, Jiang L, Mi Jeon S, Lee J, Chen Y, Liu R.