Journal ArticleDOI
Thermofluor-based high-throughput stability optimization of proteins for structural studies.
Ulrika B. Ericsson,B. Martin Hallberg,B. Martin Hallberg,George T. DeTitta,Niek Dekker,Pär Nordlund,Pär Nordlund +6 more
TLDR
A twofold increase in the number of crystallization leads was observed when the proteins were cocrystallized with stabilizing additives as compared with experiments without these additives, suggesting that thermofluor constitutes an efficient generic high-throughput method for identification of protein properties predictive of crystallizability.About:
This article is published in Analytical Biochemistry.The article was published on 2006-10-15. It has received 860 citations till now. The article focuses on the topics: Thermal shift assay.read more
Citations
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The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability
TL;DR: Differential scanning fluorimetry (DSF) is a rapid and inexpensive screening method to identify low-molecular-weight ligands that bind and stabilize purified proteins.
Journal ArticleDOI
Extrinsic fluorescent dyes as tools for protein characterization.
TL;DR: The intention of this review is to give an overview of available extrinsic dyes, explain their spectral properties, and show illustrative examples of their various applications in protein characterization.
Journal ArticleDOI
The cellular thermal shift assay for evaluating drug target interactions in cells
Rozbeh Jafari,Helena Almqvist,Hanna Axelsson,Marina Ignatushchenko,Thomas Lundbäck,Pär Nordlund,Daniel Martinez Molina +6 more
TL;DR: The cellular thermal shift assay (CETSA) allows studies of target engagement of drug candidates in a cellular context, herein exemplified with experimental data on the human kinases p38α and ERK1/2.
Journal ArticleDOI
The cap-snatching endonuclease of influenza virus polymerase resides in the PA subunit
Alexandre Dias,Denis Bouvier,Thibaut Crépin,Andrew A. McCarthy,Darren J. Hart,Florence Baudin,Stephen Cusack,Rob W.H. Ruigrok +7 more
TL;DR: In this paper, the amino-terminal 209 residues of the PA subunit contain the active site of the endonuclease active site, which is shown to be strongly activated by manganese ions, matching observations reported for the intact trimeric polymerase.
Journal ArticleDOI
Structure of mycobacterial maltokinase, the missing link in the essential GlgE-pathway
TL;DR: The structural characterization of the fourth enzyme of the pathway, mycobacterial maltokinase (Mak), is described, uncovering a eukaryotic-like kinase (ELK) fold, similar to methylthioribose kinases and aminoglycoside phosphotransferases, which has a novel N-terminal domain topologically resembling the cystatin family of protease inhibitors.
References
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Journal ArticleDOI
High-Density Miniaturized Thermal Shift Assays as a General Strategy for Drug Discovery
Michael W. Pantoliano,Eugene C. Petrella,Joseph Kwasnoski,Victor S. Lobanov,James Myslik,Edward Graf,Ted Carver,Eric D. Asel,Barry A. Springer,Pamela Lane,Francis R. Salemme +10 more
TL;DR: The general applicability of the thermal shift screening strategy was found to be an important advantage because it circumvents the need to design and retool new assays with each new therapeutic target, making it ideally suited for the quantitative high throughput drug screening and evaluation of targets derived from genomics.
Journal ArticleDOI
Evaluation of fluorescence-based thermal shift assays for hit identification in drug discovery.
Mei-Chu Lo,Ann Aulabaugh,Guixian Jin,Rebecca Cowling,Jonathan A. Bard,Michael S. Malamas,George A. Ellestad +6 more
TL;DR: It is reported that the thermal shift assay can be conducted in an inexpensive, commercially available device for temperature control and fluorescence detection and the binding affinities obtained are compared with isothermal titration calorimetry and with the IC(50) values from enzymatic assays.
Journal ArticleDOI
A deliberate approach to screening for initial crystallization conditions of biological macromolecules
Joseph R. Luft,Joseph R. Luft,Robert J. Collins,Nancy A. Fehrman,Angela Lauricella,Christina K. Veatch,George T. DeTitta,George T. DeTitta +7 more
TL;DR: Results and guidelines for the initial screening of crystallization conditions, applicable to both manual and robotic setups, are discussed and a facility that executes the rapid survey of crystallized lead conditions is described in detail.
Journal ArticleDOI
Microbatch crystallization under oil — a new technique allowing many small-volume crystallization trials
TL;DR: In this article, a computer controlled microdispenser is used to make protein crystallization samples as microbatch droplets under oil, which are then dispensed ready-mixed and with good precision.
Journal ArticleDOI
Structure of Thermotoga maritima Stationary Phase Survival Protein SurE: A Novel Acid Phosphatase
R. Zhang,Tatiana Skarina,Jonathan E. Katz,S. Beasley,A. Khachatryan,S. Vyas,Cheryl H. Arrowsmith,Steven Clarke,Aled M. Edwards,Aled M. Edwards,Andrzej Joachimiak,Alexei Savchenko +11 more
TL;DR: The structure of SurE provided information about the protein's fold, oligomeric state, and active site, but the physiologically relevant substrate(s) remains to be identified.