Kinetics of Protein-Protein Interactions at the Surface of an Optical Biosensor

1995 ◽  
Vol 231 (1) ◽  
pp. 210-217 ◽  
Author(s):  
P.R. Edwards ◽  
A. Gill ◽  
D.V. Pollardknight ◽  
M. Hoare ◽  
P.E. Buckle ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Optical Biosensor ◽  
Protein Protein Interactions ◽  
Kinetics Of

scholarly journals The Effect of Proline cis-trans Isomerization on the Folding of the C-Terminal SH2 Domain from p85

2019 ◽  
Vol 21 (1) ◽  
pp. 125
Author(s):  
Francesca Troilo ◽  
Francesca Malagrinò ◽  
Lorenzo Visconti ◽  
Angelo Toto ◽  
Stefano Gianni
Keyword(s):  
Protein Interactions ◽  
Specific Interaction ◽  
Sh2 Domain ◽  
Regulatory Subunit ◽  
Wild Type ◽  
Protein Protein Interactions ◽  
Sh2 Domains ◽  
Trans Isomerization ◽  
A Site ◽  
Kinetics Of

SH2 domains are protein domains that modulate protein–protein interactions through a specific interaction with sequences containing phosphorylated tyrosines. In this work, we analyze the folding pathway of the C-terminal SH2 domain of the p85 regulatory subunit of the protein PI3K, which presents a proline residue in a cis configuration in the loop between the βE and βF strands. By employing single and double jump folding and unfolding experiments, we demonstrate the presence of an on-pathway intermediate that transiently accumulates during (un)folding. By comparing the kinetics of folding of the wild-type protein to that of a site-directed variant of C-SH2 in which the proline was replaced with an alanine, we demonstrate that this intermediate is dictated by the peptidyl prolyl cis-trans isomerization. The results are discussed in the light of previous work on the effect of peptidyl prolyl cis-trans isomerization on folding events.

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

scholarly journals Quantifying the monomer-dimer equilibrium of tubulin with mass photometry

2020 ◽  
Author(s):  
Adam Fineberg ◽  
Thomas Surrey ◽  
Philipp Kukura
Keyword(s):  
Dissociation Constant ◽  
Protein Interactions ◽  
Building Block ◽  
Dissociation Rate ◽  
Protein Protein Interactions ◽  
Cellular Processes ◽  
Fundamental Building Block ◽  
Kinetics Of

AbstractThe αβ-tubulin heterodimer is the fundamental building block of microtubules, making it central to several cellular processes. Despite the apparent simplicity of heterodimerisation, the associated energetics and kinetics remain disputed, largely due to experimental challenges associated with quantifying affinities in the <μM range. We use mass photometry to observe tubulin monomers and heterodimers in solution simultaneously, thereby quantifying the αβ-tubulin dissociation constant (8.48±1.22 nM) and its tightening in the presence of GTP (3.69±0.65 nM), at a dissociation rate >10−2 s−1. Our results demonstrate the capabilities of mass photometry for quantifying protein-protein interactions and clarify the energetics and kinetics of tubulin heterodimerisation.Abstract Figure

The role of nucleoside triphosphate hydrolysis in transducing systems: p21ras and muscle

1992 ◽  
Vol 336 (1276) ◽  
pp. 19-24 ◽  
Keyword(s):  
Protein Interactions ◽  
Inorganic Phosphate ◽  
Signal Transduction Pathway ◽  
Basic Mechanism ◽  
Nucleoside Triphosphate ◽  
Protein Protein Interactions ◽  
Phosphate Release ◽  
Kinetics Of ◽  
Degree Of Similarity

A variety of systems use nucleoside triphosphate hydrolysis to control or provide energy for biological processes, mediated through protein-protein interactions. The nature of this coupling may vary, but often there is a degree of similarity. In this paper, two systems are compared: actomyosin in muscle and p21ras in a signal transduction pathway as yet undefined. The mechanism of the nucleotide triphosphate hydrolysis and the consequent changes in the protein-nucleotide complex have been investigated, to understand how the coupling to biological function is achieved. The basal nucleoside triphosphatase mechanisms are compared and the roles of proteins that activate the hydrolysis, actin and GAP, are discussed. The cleavage process was probed by stereochemical techniques to determine the basic mechanism, of either a phosphorylated enzyme intermediate or direct displacement of nucleoside diphosphate by water. Phosphate-water oxygen exchange probes were used to investigate nucleoside triphosphate and inorganic phosphate release steps. A new method of probing the kinetics of inorganic phosphate release directly has been developed. In muscle, this process seems likely to be related directly to force generation. In the GAP-ras system, measurement of phosphate release is allowing the mechanism of the GAP-p21ras interaction to be probed.

Molecular Recognition in the P450cam Monooxygenase System: Direct Monitoring of Protein–Protein Interactions by Using Optical Biosensor

2001 ◽  
Vol 391 (2) ◽  
pp. 255-264 ◽  
Author(s):  
Yuri D. Ivanov ◽  
Irina P. Kanaeva ◽  
Irina I. Karuzina ◽  
Alexander I. Archakov ◽  
Gaston Hui Bon Hoa ◽  
...  
Keyword(s):  
Molecular Recognition ◽  
Protein Interactions ◽  
Optical Biosensor ◽  
Monooxygenase System ◽  
Protein Protein Interactions

Kinetics of Protein-Protein Interactions of Connexins: Use of Enzyme Linked Sorbent Assays

2003 ◽  
Vol 10 (4-6) ◽  
pp. 207-210 ◽  
Author(s):  
Heather S. Duffy ◽  
Phyllis O'Donnell ◽  
Wanda Coombs ◽  
Steven M. Taffet ◽  
Mario Delmar ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Protein Protein Interactions ◽  
Kinetics Of

scholarly journals The effect of isatin on protein-protein interactions between cytochrome b5 and cytochromes P450

2017 ◽  
Vol 63 (2) ◽  
pp. 170-175 ◽  
Author(s):  
P.V. Ershov ◽  
E.O. Yablokov ◽  
Yu.V. Mezentsev ◽  
L.A. Kalushskiy ◽  
A.V. Florinskaya ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Contact Region ◽  
Cytochromes P450 ◽  
Cytochrome B5 ◽  
Optical Biosensor ◽  
Test System ◽  
Feedback Mechanism ◽  
Dissociation Rate ◽  
Protein Protein Interactions ◽  
High Concentration

Cytochromes P450 (CYP) are involved in numerous biochemical processes including metabolism of xenobiotics, biosynthesis of cholesterol, steroid hormones etc. Since some CYP catalyze indol oxidation to isatin, we have hypothesized that isatin can regulate protein-protein interactions (PPI) between components of the CYP system thus representing a (negative?) feedback mechanism. The aim of this study was to investigate a possible effect of isatin on interaction of human CYP with cytochrome b5 (CYB5A). Using the optical biosensor test system employing surface plasmon resonance (SPR) we have investigated interaction of immobilized CYB5A with various CYP in the absence and in the presence of isatin. The SPR-based experiments have shown that a high concentration of isatin (270 mM) increases Kd values for complexes CYB5A/CYP3А5 and CYB5A/CYP3A4 (twofold and threefold, respectively), but has no influence on complex formation between CYB5A and other CYP (including indol-metabolizing CYP2C19 and CYP2E1). Isatin injection to the optical biosensor chip with the preformed molecular complex CYB5A/CYP3A4 caused a 30%-increase in its dissociation rate. Molecular docking manipulations have shown that isatin can influence interaction of CYP3А5 or CYP3A4 with CYB5A acting at the contact region of CYB5A/CYP.

Effect of hydrogel matrix on binding kinetics of protein–protein interactions on sensor surface

2002 ◽  
Vol 456 (2) ◽  
pp. 201-208 ◽  
Author(s):  
Chi-Chun Fong ◽  
Man-Sau Wong ◽  
Wang-Fun Fong ◽  
Mengsu Yang
Keyword(s):  
Protein Interactions ◽  
Binding Kinetics ◽  
Sensor Surface ◽  
Protein Protein Interactions ◽  
Hydrogel Matrix ◽  
Kinetics Of
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