scholarly journals High-Throughput Micro-Characterization of RNA–Protein Interactions

2019 ◽  
pp. 519-531
Author(s):  
Sara Gómez ◽  
Francisco J. Fernández ◽  
M. Cristina Vega
Keyword(s):  
High Throughput ◽  
Protein Interactions

scholarly journals High-throughput characterization of protein–protein interactions by reprogramming yeast mating

2017 ◽  
Vol 114 (46) ◽  
pp. 12166-12171 ◽  
Author(s):  
David Younger ◽  
Stephanie Berger ◽  
David Baker ◽  
Eric Klavins
Keyword(s):  
High Throughput ◽  
Protein Interaction ◽  
Protein Interactions ◽  
Binding Proteins ◽  
Protein Interaction Networks ◽  
Interaction Networks ◽  
Protein Protein Interactions ◽  
Mating Efficiency ◽  
Extracellular Environment

High-throughput methods for screening protein–protein interactions enable the rapid characterization of engineered binding proteins and interaction networks. While existing approaches are powerful, none allow quantitative library-on-library characterization of protein interactions in a modifiable extracellular environment. Here, we show that sexual agglutination ofSaccharomyces cerevisiaecan be reprogrammed to link interaction strength with mating efficiency using synthetic agglutination (SynAg). Validation of SynAg with 89 previously characterized interactions shows a log-linear relationship between mating efficiency and protein binding strength for interactions withKds ranging from below 500 pM to above 300 μM. Using induced chromosomal translocation to pair barcodes representing binding proteins, thousands of distinct interactions can be screened in a single pot. We demonstrate the ability to characterize protein interaction networks in a modifiable environment by introducing a soluble peptide that selectively disrupts a subset of interactions in a representative network by up to 800-fold. SynAg enables the high-throughput, quantitative characterization of protein–protein interaction networks in a fully defined extracellular environment at a library-on-library scale.

scholarly journals High-Throughput Characterization of Protein-Protein Interactions by Reprogramming Yeast Mating

2017 ◽  
Author(s):  
David Younger ◽  
Stephanie Berger ◽  
David Baker ◽  
Eric Klavins
Keyword(s):  
High Throughput ◽  
Protein Interactions ◽  
Interaction Strength ◽  
Protein Interaction Networks ◽  
Interaction Networks ◽  
Protein Protein Interactions ◽  
Quantitative Characterization ◽  
Mating Efficiency ◽  
Extracellular Environment

AbstractHigh-throughput methods for screening protein-protein interactions enable the rapid characterization of engineered binding proteins and interaction networks. While existing approaches are powerful, none allow quantitative library-on-library characterization of protein interactions in a modifiable extracellular environment. Here, we show that sexual agglutination of S. cerevisiae can be reprogrammed to link interaction strength with mating efficiency using synthetic agglutination (SynAg). Validation of SynAg with 89 previously characterized interactions shows a log-linear relationship between mating efficiency and protein binding strength for interactions with KD’s ranging from below 500 pM to above 300 μM. Using induced chromosomal translocation to pair barcodes representing binding proteins, thousands of distinct interactions can be screened in a single pot. We demonstrate the ability to characterize protein interaction networks in a modifiable environment by introducing a soluble peptide that selectively disrupts a subset of interactions in a representative network by up to 800-fold. SynAg enables the high-throughput, quantitative characterization of protein-protein interaction networks in a fully-defined extracellular environment at a library-on-library scale.Significance StatementDe novo engineering of protein binders often requires experimental screening to select functional variants from a design library. We have achieved high-throughput, quantitative characterization of protein-protein binding interactions without requiring purified recombinant proteins, by linking interaction strength with yeast mating. Using a next-generation sequencing output, we have characterized protein networks consisting of thousands of pairwise interactions in a single tube and have demonstrated the effect of changing the binding environment. This approach addresses an existing bottleneck in protein binder design by enabling the high-throughput and quantitative characterization of binding strength between designed protein libraries and multiple target proteins in a fully defined environment.

Combinatorial Fabrication and High-Throughput Characterization of Thin Film Metal Oxide Libraries for Solar water Splitting

2018 ◽  
Author(s):  
Alfred Ludwig ◽  
Mona Nowak ◽  
Swati Kumari ◽  
Helge S. Stein ◽  
Ramona Gutkowski ◽  
...  
Keyword(s):  
Thin Film ◽  
Metal Oxide ◽  
Water Splitting ◽  
High Throughput ◽  
Solar Water ◽  
Solar Water Splitting

scholarly journals Database of ab initio L-edge X-ray absorption near edge structure

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Yiming Chen ◽  
Chi Chen ◽  
Chen Zheng ◽  
Shyam Dwaraknath ◽  
Matthew K. Horton ◽  
...  
Keyword(s):  
High Throughput ◽  
Scattering Theory ◽  
Research Community ◽  
Metal Compounds ◽  
Edge Structure ◽  
X Ray ◽  
Initial Release ◽  
Computational Workflow ◽  
X Ray Absorption

AbstractThe L-edge X-ray Absorption Near Edge Structure (XANES) is widely used in the characterization of transition metal compounds. Here, we report the development of a database of computed L-edge XANES using the multiple scattering theory-based FEFF9 code. The initial release of the database contains more than 140,000 L-edge spectra for more than 22,000 structures generated using a high-throughput computational workflow. The data is disseminated through the Materials Project and addresses a critical need for L-edge XANES spectra among the research community.

High-Throughput Synthesis and Characterization of a Combinatorial Materials Library in Bulk Alloys

2021 ◽  
Vol 52 (4) ◽  
pp. 1159-1168
Author(s):  
Lei Zhao ◽  
Yuanxun Zhou ◽  
Hui Wang ◽  
Xuebin Chen ◽  
Lixia Yang ◽  
...  
Keyword(s):  
High Throughput ◽  
Synthesis And Characterization ◽  
Combinatorial Materials ◽  
Bulk Alloys

High-Throughput Screening and Characterization of a High-Density Soybean Mutant Library Elucidate the Biosynthesis Pathway of Triterpenoid Saponins

2019 ◽  
Vol 60 (5) ◽  
pp. 1082-1097 ◽  
Author(s):  
Panneerselvam Krishnamurthy ◽  
Yukiko Fujisawa ◽  
Yuya Takahashi ◽  
Hanako Abe ◽  
Kentaro Yamane ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
High Density ◽  
Biosynthesis Pathway ◽  
Mutant Library ◽  
Triterpenoid Saponins

scholarly journals Size-based characterization of adalimumab and TNF-α interactions using flow induced dispersion analysis: assessment of avidity-stabilized multiple bound species

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Morten E. Pedersen ◽  
Ragna M. S. Haegebaert ◽  
Jesper Østergaard ◽  
Henrik Jensen
Keyword(s):  
Protein Interactions ◽  
Free Ligand ◽  
Local Environment ◽  
Dispersion Analysis ◽  
Surface Immobilization ◽  
Covalent Bonds ◽  
Essential Information ◽  
Tnf Α

AbstractThe understanding and characterization of protein interactions is crucial for elucidation of complicated biomolecular processes as well as for the development of new biopharmaceutical therapies. Often, protein interactions involve multiple binding, avidity, oligomerization, and are dependent on the local environment. Current analytical methodologies are unable to provide a detailed mechanistic characterization considering all these parameters, since they often rely on surface immobilization, cannot measure under biorelevant conditions, or do not feature a structurally-related readout for indicating formation of multiple bound species. In this work, we report the use of flow induced dispersion analysis (FIDA) for in-solution characterization of complex protein interactions under in vivo like conditions. FIDA is an immobilization-free ligand binding methodology employing Taylor dispersion analysis for measuring the hydrodynamic radius (size) of biomolecular complexes. Here, the FIDA technology is utilized for a size-based characterization of the interaction between TNF-α and adalimumab. We report concentration-dependent complex sizes, binding affinities (Kd), kinetics, and higher order stoichiometries, thus providing essential information on the TNF-α–adalimumab binding mechanism. Furthermore, it is shown that the avidity stabilized complexes involving formation of multiple non-covalent bonds are formed on a longer timescale than the primary complexes formed in a simple 1 to 1 binding event.

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