Quantitative Real-Time Imaging of Protein–Protein Interactions by LSPR Detection with Micropatterned Gold Nanoparticles

2013 ◽  
Vol 85 (20) ◽  
pp. 9564-9571 ◽  
Author(s):  
Maniraj Bhagawati ◽  
Changjiang You ◽  
Jacob Piehler
Keyword(s):  
Gold Nanoparticles ◽  
Real Time ◽  
Protein Interactions ◽  
Protein Protein Interactions ◽  
Real Time Imaging

scholarly journals High-Sensitivity Real-Time Imaging of Dual Protein-Protein Interactions in Living Subjects Using Multicolor Luciferases

PLoS ONE ◽  
2009 ◽  
Vol 4 (6) ◽  
pp. e5868 ◽  
Author(s):  
Naoki Hida ◽  
Muhammad Awais ◽  
Masaki Takeuchi ◽  
Naoto Ueno ◽  
Mayuri Tashiro ◽  
...  
Keyword(s):  
Real Time ◽  
Protein Interactions ◽  
High Sensitivity ◽  
Protein Protein Interactions ◽  
Real Time Imaging

scholarly journals EndoBind detects endogenous protein-protein interactions in real time

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Anke Bill ◽  
Sheryll Espinola ◽  
Daniel Guthy ◽  
Jacob R. Haling ◽  
Mylene Lanter ◽  
...  
Keyword(s):  
Real Time ◽  
High Throughput ◽  
Protein Interactions ◽  
Continuous Monitoring ◽  
Protein Protein Interactions ◽  
Dimer Formation ◽  
Endogenous Protein ◽  
Extended Duration ◽  
Substrate Addition

AbstractWe present two high-throughput compatible methods to detect the interaction of ectopically expressed (RT-Bind) or endogenously tagged (EndoBind) proteins of interest. Both approaches provide temporal evaluation of dimer formation over an extended duration. Using examples of the Nrf2-KEAP1 and the CRAF-KRAS-G12V interaction, we demonstrate that our method allows for the detection of signal for more than 2 days after substrate addition, allowing for continuous monitoring of endogenous protein-protein interactions in real time.

Distance Controlled and Electrically Driven Photoluminescence Quench From Quantum Dot-Au Complexes

2010 ◽  
Vol 1257 ◽  
Author(s):  
Zhitao Kang ◽  
Jie Xu ◽  
Dinal Andreasen ◽  
Brent Karl Wagner
Keyword(s):  
Gold Nanoparticles ◽  
Protein Interactions ◽  
Near Infrared ◽  
Incident Light ◽  
Gold Surface ◽  
Electric Signal ◽  
Protein Protein Interactions ◽  
Quenching Effect ◽  
Quenching Efficiency ◽  
Pl Quenching

AbstractQuantum Dots (QDs) bound to gold nanoparticles have shown photoluminescence (PL) quenching dependent on distance between the two particles. The incident light from the QD couples to plasmon excitation of the metal when the frequencies of the light and the surface plasmon resonance (SPR) coincide, leading to a reduction in emitted PL in the system. The quenching effect of gold nanoparticles on QDs was used to study protein-protein interactions with the potential for drug screening applications. CdTe and CdHgTe QDs with emission wavelengths from 500˜900nm were synthesized and gold nanospheres and nanorods with controlled absorption in the visible and near-infrared (NIR) wavelength regions were prepared. The PL quenching of QD-Protein-Protein-Au complexes was studied as a function of Au concentration, QD size and protein type. A quenching efficiency of up to 90% was observed. The QD-Au complexes were also studied for electric potential sensing. The surface of the QDs was negatively charged due to thiol ligands capping. By applying a positive potential on the gold or gold nanoparticle attached substrate, the local electric field between the substrate and the statically charged QDs would pull the QDs closer to the gold surface and quench the QD PL. PL quenching of QD with Au was studied as a function of electric signal and QD type. In this methodology, electric signals were effectively converted to optical signals.

scholarly journals DNA-mediated engineering of multicomponent enzyme crystals

2015 ◽  
Vol 112 (15) ◽  
pp. 4564-4569 ◽  
Author(s):  
Jeffrey D. Brodin ◽  
Evelyn Auyeung ◽  
Chad A. Mirkin
Keyword(s):  
Gold Nanoparticles ◽  
Protein Interactions ◽  
Building Blocks ◽  
Inorganic Nanoparticles ◽  
Protein Protein Interactions ◽  
Dna Interactions ◽  
Interparticle Interactions ◽  
Crystalline Materials ◽  
Catalytically Active ◽  
Surface Chemistries

The ability to predictably control the coassembly of multiple nanoscale building blocks, especially those with disparate chemical and physical properties such as biomolecules and inorganic nanoparticles, has far-reaching implications in catalysis, sensing, and photonics, but a generalizable strategy for engineering specific contacts between these particles is an outstanding challenge. This is especially true in the case of proteins, where the types of possible interparticle interactions are numerous, diverse, and complex. Herein, we explore the concept of trading protein–protein interactions for DNA–DNA interactions to direct the assembly of two nucleic-acid–functionalized proteins with distinct surface chemistries into six unique lattices composed of catalytically active proteins, or of a combination of proteins and DNA-modified gold nanoparticles. The programmable nature of DNA–DNA interactions used in this strategy allows us to control the lattice symmetries and unit cell constants, as well as the compositions and habit, of the resulting crystals. This study provides a potentially generalizable strategy for constructing a unique class of materials that take advantage of the diverse morphologies, surface chemistries, and functionalities of proteins for assembling functional crystalline materials.

scholarly journals QCM Biosensor Based on Polydopamine Surface for Real-Time Analysis of the Binding Kinetics of Protein-Protein Interactions

Polymers ◽  
2017 ◽  
Vol 9 (12) ◽  
pp. 482 ◽  
Author(s):  
Chunli Wu ◽  
Xueming Li ◽  
Siyu Song ◽  
Yuxin Pei ◽  
Lili Guo ◽  
...  
Keyword(s):  
Real Time ◽  
Protein Interactions ◽  
Binding Kinetics ◽  
Protein Protein Interactions ◽  
Time Analysis ◽  
Real Time Analysis ◽  
Kinetics Of

Cancer-targeted functional gold nanoparticles for apoptosis induction and real-time imaging based on FRET

Nanoscale ◽  
2014 ◽  
Vol 6 (16) ◽  
pp. 9531 ◽  
Author(s):  
Wei-Hai Chen ◽  
Guo-Feng Luo ◽  
Xiao-Ding Xu ◽  
Hui-Zhen Jia ◽  
Qi Lei ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
Real Time ◽  
Apoptosis Induction ◽  
Real Time Imaging

scholarly journals The inner mechanics of Rhodopsin Guanylyl Cyclase during 2cGMP-formation revealed by real-time FTIR spectroscopy

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Paul Fischer ◽  
Shatanik Mukherjee ◽  
Enrico Peter ◽  
Matthias Broser ◽  
Franz Bartl ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Real Time ◽  
Protein Interactions ◽  
Uv Light ◽  
Spectroscopic Characterization ◽  
Binding Pocket ◽  
Molecular Signaling ◽  
Protein Protein Interactions ◽  
Regulatory Influence ◽  
Truncated Variants

Enzymerhodopsins represent a recently discovered class of rhodopsins which includes histidine kinase rhodopsin, rhodopsin phosphodiesterases and rhodopsin guanylyl cyclases (RGCs). The regulatory influence of the rhodopsin domain on the enzyme activity is only partially understood and holds the key for a deeper understanding of intra-molecular signaling pathways. Here we present a UV-Vis and FTIR study about the light-induced dynamics of a RGC from the fungus Catenaria anguillulae, which provides insights into the catalytic process. After the spectroscopic characterization of the late rhodopsin photoproducts, we analyzed truncated variants and revealed the involvement of the cytosolic N-terminus in the structural rearrangements upon photo-activation of the protein. We tracked the catalytic reaction of RGC and the free GC domain independently by UV-light induced release of GTP from the photolabile NPE-GTP substrate. Our results show substrate binding to the dark-adapted RGC and GC alike and reveal differences between the constructs attributable to the regulatory influence of the rhodopsin on the conformation of the binding pocket. By monitoring the phosphate rearrangement during cGMP and pyrophosphate formation in light-activated RGC, we were able to confirm the M state as the active state of the protein. The described setup and experimental design enable real-time monitoring of substrate turnover in light-activated enzymes on a molecular scale, thus opening the pathway to a deeper understanding of enzyme activity and protein-protein interactions.

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