Using Fluorescence Anisotropy for Ligand Binding Kinetics of Membrane Proteins

2018 ◽  
Vol 93 (1) ◽  
pp. e63 ◽  
Author(s):  
Kirsten N. Swonger ◽  
Anne S. Robinson
Keyword(s):  
Membrane Proteins ◽  
Ligand Binding ◽  
Fluorescence Anisotropy ◽  
Binding Kinetics ◽  
Kinetics Of

scholarly journals Linking 3D and 2D binding kinetics of membrane proteins by multiscale simulations

2014 ◽  
Vol 23 (12) ◽  
pp. 1789-1799 ◽  
Author(s):  
Zhong-Ru Xie ◽  
Jiawen Chen ◽  
Yinghao Wu
Keyword(s):  
Membrane Proteins ◽  
Binding Kinetics ◽  
Multiscale Simulations ◽  
Kinetics Of

scholarly journals Spectroscopic analyses of the binding kinetics of 15d-PGJ2 to the PPARγ ligand-binding domain by multi-wavelength global fitting

2006 ◽  
Vol 393 (3) ◽  
pp. 749-755 ◽  
Author(s):  
Takuma Shiraki ◽  
Takashi S. Kodama ◽  
Sayaka Shiki ◽  
Tatsuo Nakagawa ◽  
Hisato Jingami
Keyword(s):  
Ligand Binding ◽  
Intermediate State ◽  
Covalent Binding ◽  
Binding Domain ◽  
Ligand Binding Domain ◽  
Binding Kinetics ◽  
Multi Wavelength ◽  
Lipid Metabolites ◽  
Kinetics Of ◽  
Global Fitting

PPARγ (peroxisome proliferator-activated receptor γ) is a nuclear receptor that is activated by natural lipid metabolites, including 15d-PGJ2 (15-deoxy-Δ12,14-prostaglandin J2). We previously reported that several oxidized lipid metabolites covalently bind to PPARγ through a Michael-addition to activate transcription. To separate the ligand-entering (dock) and covalent-binding (lock) steps in PPARγ activation, we investigated the binding kinetics of 15d-PGJ2 to the PPARγ LBD (ligand-binding domain) by stopped-flow spectroscopy. We analysed the spectral changes of 15d-PGJ2 by multi-wavelength global fitting based on a two-step chemical reaction model, in which an intermediate state represents the 15d-PGJ2–PPARγ complex without covalent binding. The extracted spectrum of the intermediate state in wild-type PPARγ was quite similar to the observed spectrum of 15d-PGJ2 in the C285S mutant, which cannot be activated by 15d-PGJ2, indicating that the complex remains in the inactive, intermediate state in the mutant. Thus ‘lock’ rather than ‘dock’ is one of the critical steps in PPARγ activation by 15d-PGJ2.

scholarly journals Functional consequences of haem orientational disorder in sperm-whale and yellow-fin-tuna myoglobins

1987 ◽  
Vol 243 (1) ◽  
pp. 205-210 ◽  
Author(s):  
H S Aojula ◽  
M T Wilson ◽  
I E G Morrison
Keyword(s):  
Ligand Binding ◽  
Kinetic Parameters ◽  
Sperm Whale ◽  
Binding Kinetics ◽  
Orientational Disorder ◽  
Binding Properties ◽  
Functional Consequences ◽  
Kinetics Of ◽  
Sperm Whale Myoglobin ◽  
O2 Dissociation

Ligand-binding kinetics of native and reconstituted sperm-whale myoglobin were studied in relation to haem orientational disorder by rapid kinetic methods. In addition, native yellow-fin-tuna myoglobin with significant amount of haem disorder was also used. The O2 dissociation and association rates were found for the proteins with different degrees of haem disorder, and these results suggest that the isomers are characterized by almost identical kinetic parameters. Rates of CO recombination after photolysis were also identical for the two orientational isomers. The results clearly indicate that the rotation of the haem about the alpha-gamma meso axis has little or no effect on the ligand-binding properties of these myoglobins.

Ligand Binding Kinetics of the Quorum Sensing Regulator PqsR

Biochemistry ◽  
2013 ◽  
Vol 52 (25) ◽  
pp. 4433-4438 ◽  
Author(s):  
Martin Welch ◽  
James T. Hodgkinson ◽  
Jeremy Gross ◽  
David R. Spring ◽  
Thomas Sams
Keyword(s):  
Quorum Sensing ◽  
Ligand Binding ◽  
Binding Kinetics ◽  
Kinetics Of

scholarly journals Characterization of the Heme Pocket Structure and Ligand Binding Kinetics of Non-symbiotic Hemoglobins from the Model Legume Lotus japonicus

2017 ◽  
Vol 8 ◽  
Author(s):  
Laura Calvo-Begueria ◽  
Bert Cuypers ◽  
Sabine Van Doorslaer ◽  
Stefania Abbruzzetti ◽  
Stefano Bruno ◽  
...  
Keyword(s):  
Ligand Binding ◽  
Lotus Japonicus ◽  
Binding Kinetics ◽  
Model Legume ◽  
Heme Pocket ◽  
Kinetics Of

scholarly journals Label-free measuring and mapping of binding kinetics of membrane proteins in single living cells

2012 ◽  
Vol 4 (10) ◽  
pp. 846-853 ◽  
Author(s):  
Wei Wang ◽  
Yunze Yang ◽  
Shaopeng Wang ◽  
Vinay J. Nagaraj ◽  
Qiang Liu ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Living Cells ◽  
Binding Kinetics ◽  
Label Free ◽  
Single Living Cells ◽  
Kinetics Of ◽  
Single Living

scholarly journals Investigations of Low-Frequency Vibrational Dynamics and Ligand Binding Kinetics of Cystathionine β-Synthase

2010 ◽  
Vol 114 (9) ◽  
pp. 3294-3306 ◽  
Author(s):  
Venugopal Karunakaran ◽  
Abdelkrim Benabbas ◽  
Yuhan Sun ◽  
Zhenyu Zhang ◽  
Sangita Singh ◽  
...  
Keyword(s):  
Ligand Binding ◽  
Low Frequency ◽  
Binding Kinetics ◽  
Vibrational Dynamics ◽  
Kinetics Of
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