Time-Resolved Tryptophan Fluorescence of Fragment 1-86 of Factor X and the Influence of Membrane Binding

[img]PDF - Authorized users only
Language: English
176Kb
Title:Time-Resolved Tryptophan Fluorescence of Fragment 1-86 of Factor X and the Influence of Membrane Binding
Creators:
Häfner, Angelika
Beneš, Martin
Mérola, Fabienne
Duportail, Guy
Schneider, Friedemann W.
Hof, Martin
Journal or Publication Title:
Collection of Czechoslovak Chemical Communications, 67, 12, pp. 1872-1882
Uncontrolled Keywords:EGF-like domain, Enzymes, Blood coagulation, Factor Xa, Protein fluorescence, Synchrotron radiation, Lipid-specificity, Prothrombinase, Gla domain

Abstract

Membrane binding of the N-terminus of the prothrombinase reaction enzyme, fragment 1-86 of factor X (FX F1-86), to phospholipid vesicles was studied using time-resolved fluorescence spectroscopy of the tryptophan residue Trp41 which is located in the membrane binding part of this protein (Gla domain). Fluorescence lifetimes were determined by the time-correlated single photon counting technique using synchrotron radiation as the excitation source. Different negatively charged lipid vesicles containing either phosphatidylserine or phosphatidylglycerol mixed with phosphatidylcholine were investigated. Pure synthetic lipids (dilauroyl and dioleoyl lipids) as well as natural lipids (from egg yolk or bovine brain) were used in membrane binding studies. Multiexponential data analysis supports the results already found for FX F1-86 in the presence of calcium ions. For all lipid systems we obtained decay times in the range of 0.2, 0.6, 2.6, 5.6 ns. Moreover, a long-lifetime component with minor contribution to the decay amplitude (&lt;5%) was found. The fluorescence lifetimes do not show any wavelength dependence. There was no change in the fluorescence decay times upon membrane binding, which might indicate that a lipid-specific conformational change in the Gla domain of factor X does not take place upon membrane binding. <p>

Title:Time-Resolved Tryptophan Fluorescence of Fragment 1-86 of Factor X and the Influence of Membrane Binding
Creators:
Häfner, Angelika
Beneš, Martin
Mérola, Fabienne
Duportail, Guy
Schneider, Friedemann W.
Hof, Martin
Uncontrolled Keywords:EGF-like domain, Enzymes, Blood coagulation, Factor Xa, Protein fluorescence, Synchrotron radiation, Lipid-specificity, Prothrombinase, Gla domain
Divisions:Life and Chemical Sciences > Institute of Organic Chemistry and Biochemistry > Collection of Czechoslovak Chemical Communications
Journal or Publication Title:Collection of Czechoslovak Chemical Communications
Volume:67
Number:12
Page Range:pp. 1872-1882
ISSN:0010-0765
E-ISSN:1212-6950
Publisher:Institute of Organic Chemistry and Biochemistry
Related URLs:
URLURL Type
http://dx.doi.org/10.1135/cccc20021872UNSPECIFIED
ID Code:2039
Item Type:Article
Deposited On:06 Feb 2009 17:15
Last Modified:06 Feb 2009 16:15

Citation

Häfner, Angelika; Beneš, Martin; Mérola, Fabienne; Duportail, Guy; Schneider, Friedemann W.; Hof, Martin (2002) Time-Resolved Tryptophan Fluorescence of Fragment 1-86 of Factor X and the Influence of Membrane Binding. Collection of Czechoslovak Chemical Communications, 67 (12). pp. 1872-1882. ISSN 0010-0765

Repository Staff Only: item control page