Human factor VIIa and its complex with soluble tissue factor: evaluation of asymmetry and conformational dynamics by ultracentrifugation and fluorescence anisotropy decay methods

Biochemistry ◽  
1993 ◽  
Vol 32 (12) ◽  
pp. 3005-3012 ◽  
Author(s):  
Evan Waxman ◽  
William R. Laws ◽  
Thomas M. Laue ◽  
Yale Nemerson ◽  
J. B. Alexander Ross
Keyword(s):  
Tissue Factor ◽  
Fluorescence Anisotropy ◽  
Human Factor ◽  
Factor Viia ◽  
Conformational Dynamics ◽  
Fluorescence Anisotropy Decay ◽  
Anisotropy Decay ◽  
Factor Evaluation

Activation of Factor X by Factor VIIa Complexed with Human-mouse Tissue Factor Chimeras Requires Human Exon 3

1996 ◽  
Vol 76 (03) ◽  
pp. 361-368 ◽  
Author(s):  
Carrie H Fang ◽  
T-C Lin ◽  
Arabinda Guha ◽  
Yale Nemerson ◽  
William H Konigsberg
Keyword(s):  
Tissue Factor ◽  
Human Factor ◽  
Factor Viia ◽  
Mouse Tissue ◽  
Factor X ◽  
Lower Affinity ◽  
E Coli ◽  
Sequence Elements ◽  
Specific Activities ◽  
Human And Mouse

SummaryIn an attempt to define sequence elements in human and mouse tissue factor (TF) that are responsible for the species specificity observed in their interaction with human factor VIIa (HVIIa), we constructed human-mouse chimeric TF cDNAs, inserted them into plasmid vectors, and induced their expression in E.coli. Assays for procoagulant activity were carried out with the resulting E. coli lysates using (HVIIa) human and mouse (MVIIa). The ratio of the procoagulant activities, HVIIa/MVIIa, revealed that human TF exon 3 was essential for activity when the TF:VIIa complex was formed with HVIIa. By ligating the maltose binding protein (MBP) gene to TF cDNAs it was possible to construct, express and purify MBP-TF chimeras as well as to estimate their specific activities. With selected MBP-TF chimeras and HVIIa we determined kinetic parameters for the activation of human factor X. Replacement of exon 3 in human TF cDNA with the corresponding exon from mouse TF cDNA resulted in both lower affinity for HVIIa and failure to convert bound HVIIa into a potent protease

Investigation of the mobility of polybutadienes: 2. Fluorescence anisotropy decay of rod-like rigid probes

Polymer ◽  
1989 ◽  
Vol 30 (1) ◽  
pp. 51-57 ◽  
Author(s):  
Mamadou Fofana ◽  
Valérie Veissier ◽  
Jean Louis Viovy ◽  
Lucien Monnerie
Keyword(s):  
Fluorescence Anisotropy ◽  
Fluorescence Anisotropy Decay ◽  
Anisotropy Decay

scholarly journals A 384-HTS for Human Factor VIIa: Comparison With 96- and 864-Well Formats

1997 ◽  
Vol 2 (3) ◽  
pp. 171-178 ◽  
Author(s):  
John C.W. Comley ◽  
Alastair Binnie ◽  
Caroline Bonk ◽  
John G. Houston
Keyword(s):  
Tissue Factor ◽  
Dose Response ◽  
Human Factor ◽  
Factor Viia ◽  
Assay Development ◽  
Fluorescent Detection ◽  
Response Curves ◽  
Liquid Handling ◽  
Dose Response Curves ◽  
Assay Volume

A homogenous fluorescent HTS for recombinant human factor VIIa (FVIIa) using soluble tissue factor has been developed in 384-well microplates. In this report we discuss our experiences with assay development, liquid handling using a Tomtec Quadra and Matrix PlateMate, fluorescent detection and screening of -200,000 compounds against FVIIa in 384-well plate format. Assays using the entire Helix 864-well plate were prototyped using contact dispensing with a modified Hamilton Microlab 2200. FVIIa was used as a model assay to compare between 96-, 384-, and 864-plate formats in a total assay volume of 100, 25, and 10 μl, respectively. FVIIa was assayed in 864 to the same degree of sensitivity as 384- and 96-well assays and dose-response curves for a standard inhibitor (benzamidine) in the FVIIa assay were identical in all plate formats. Finally, we review the prospects for HTS in 864-well microplates.

Fluorescence anisotropy decay of ethidium bound to chromatin

1981 ◽  
Vol 13 (1) ◽  
pp. 77-87 ◽  
Author(s):  
Daniel Genest ◽  
Georgette Sabeur ◽  
Philippe Wahl ◽  
Jean-Claude Auchet
Keyword(s):  
Fluorescence Anisotropy ◽  
Fluorescence Anisotropy Decay ◽  
Anisotropy Decay

Comment on ““Where does the fluorescing moiety reside in a carbon dot?” – Investigations based on fluorescence anisotropy decay and resonance energy transfer dynamics” by A. Das, D. Roy, C. K. De and P. K. Mandal, Phys. Chem. Chem. Phys., 2018, 20, 2251

2019 ◽  
Vol 21 (24) ◽  
pp. 13368-13369 ◽  
Author(s):  
Hem C. Joshi
Keyword(s):  
Physical Chemistry ◽  
Energy Transfer ◽  
Fluorescence Anisotropy ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Chem Phys ◽  
Chemical Physics ◽  
Fluorescence Anisotropy Decay ◽  
Anisotropy Decay ◽  
Phys Chem Chem Phys

In a recent paper published in Physical Chemistry Chemical Physics (Phys. Chem. Chem. Phys., 2018, 20, 2251–2259), Förster resonance energy transfer (FRET) between carbon dots and rhodamine 123 has been reported.

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