Probing the binding kinetics of proinflammatory cytokine–antibody interactions using dual color fluorescence cross correlation spectroscopy

The Analyst ◽  
2011 ◽  
Vol 136 (10) ◽  
pp. 2111 ◽  
Author(s):  
Chia-Yan Wu ◽  
Chuan-Keng Huang ◽  
Chao-Yu Chung ◽  
I-Ping Huang ◽  
Yeukuang Hwu ◽  
...  
Keyword(s):  
Proinflammatory Cytokine ◽  
Cross Correlation ◽  
Correlation Spectroscopy ◽  
Binding Kinetics ◽  
Dual Color ◽  
Antibody Interactions ◽  
Kinetics Of

scholarly journals Dual-Color Fluorescence Cross-Correlation Spectroscopy for Monitoring the Kinetics of Enzyme-Catalyzed Reactions

Methods ◽  
2001 ◽  
Vol 24 (2) ◽  
pp. 104-116 ◽  
Author(s):  
Markus Rarbach ◽  
Ulrich Kettling ◽  
Andre Koltermann ◽  
Manfred Eigen
Keyword(s):  
Cross Correlation ◽  
Correlation Spectroscopy ◽  
Dual Color ◽  
Catalyzed Reactions ◽  
Enzyme Catalyzed Reactions ◽  
Kinetics Of

scholarly journals Fluorescence Cross-Correlation Spectroscopy Yields True Affinity and Binding Kinetics of Plasmodium Lactate Transport Inhibitors

2021 ◽  
Vol 14 (8) ◽  
pp. 757
Author(s):  
Iga Jakobowska ◽  
Frank Becker ◽  
Stefano Minguzzi ◽  
Kerrin Hansen ◽  
Björn Henke ◽  
...  
Keyword(s):  
Rate Constants ◽  
Cross Correlation ◽  
Fluorescent Protein ◽  
Correlation Spectroscopy ◽  
Confocal Imaging ◽  
Binding Kinetics ◽  
Acceptor Site ◽  
Lactate Transport ◽  
Entry Site ◽  
Hydrogen Bond Acceptor

Blocking lactate export in the parasitic protozoan Plasmodium falciparum is a novel strategy to combat malaria. We discovered small drug-like molecules that inhibit the sole plasmodial lactate transporter, PfFNT, and kill parasites in culture. The pentafluoro-3-hydroxy-pent-2-en-1-one BH296 blocks PfFNT with nanomolar efficiency but an in vitro selected PfFNT G107S mutation confers resistance against the drug. We circumvented the mutation by introducing a nitrogen atom as a hydrogen bond acceptor site into the aromatic ring of the inhibitor yielding BH267.meta. The current PfFNT inhibitor efficiency values were derived from yeast-based lactate transport assays, yet direct affinity and binding kinetics data are missing. Here, we expressed PfFNT fused with a green fluorescent protein in human embryonic kidney cells and generated fluorescent derivatives of the inhibitors, BH296 and BH267.meta. Using confocal imaging, we confirmed the location of the proposed binding site at the cytosolic transporter entry site. We then carried out fluorescence cross-correlation spectroscopy measurements to assign true Ki-values, as well as kon and koff rate constants for inhibitor binding to PfFNT wildtype and the G107S mutant. BH296 and BH267.meta gave similar rate constants for binding to PfFNT wildtype. BH296 was inactive on PfFNT G107S, whereas BH267.meta bound the mutant protein albeit with weaker affinity than to PfFNT wildtype. Eventually, using a set of PfFNT inhibitor compounds, we found a robust correlation of the results from the biophysical FCCS binding assay to inhibition data of the functional transport assay.

Dual-color fluorescence cross-correlation spectroscopy on a planar optofluidic chip

Lab on a Chip ◽  
2011 ◽  
Vol 11 (8) ◽  
pp. 1502 ◽  
Author(s):  
A. Chen ◽  
M. M. Eberle ◽  
E. J. Lunt ◽  
S. Liu ◽  
K. Leake ◽  
...  
Keyword(s):  
Cross Correlation ◽  
Correlation Spectroscopy ◽  
Dual Color ◽  
Optofluidic Chip

A fluorescent variant of a protein from the stony coral Montipora facilitates dual-color single-laser fluorescence cross-correlation spectroscopy

2006 ◽  
Vol 24 (5) ◽  
pp. 577-581 ◽  
Author(s):  
Takako Kogure ◽  
Satoshi Karasawa ◽  
Toshio Araki ◽  
Kenta Saito ◽  
Masataka Kinjo ◽  
...  
Keyword(s):  
Cross Correlation ◽  
Correlation Spectroscopy ◽  
Laser Fluorescence ◽  
Stony Coral ◽  
Single Laser ◽  
Dual Color

Dual-color Fluorescence Cross Correlation Spectroscopy on an Integrated Optofluidic Chip

2010 ◽  
Author(s):  
A. Chen ◽  
M.I. Rudenko ◽  
E.J. Lunt ◽  
B.S. Phillips ◽  
A. R. Hawkins ◽  
...  
Keyword(s):  
Cross Correlation ◽  
Correlation Spectroscopy ◽  
Dual Color ◽  
Optofluidic Chip

Parallel dual-color fluorescence cross-correlation spectroscopy using diffractive optical elements

2005 ◽  
Vol 10 (5) ◽  
pp. 054008 ◽  
Author(s):  
Michael Gösch ◽  
Hans Blom ◽  
Sylvain Anderegg ◽  
Kerstin Korn ◽  
Per Thyberg ◽  
...  
Keyword(s):  
Cross Correlation ◽  
Correlation Spectroscopy ◽  
Diffractive Optical Elements ◽  
Optical Elements ◽  
Dual Color
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