Tyrosine fluorescence analysis of apolipophorin III-lipopolysaccharide interaction.

Archives of Biochemistry and Biophysics
Leonardo J LeonPaul M M Weers

Abstract

Apolipophorin III (apoLp-III) is an exchangeable apolipoprotein that binds to lipopolysaccharides (LPS). Polyacrylamide gel electrophoresis analysis demonstrated that apoLp-III from Galleria mellonella associated with various truncated LPS variants, including lipid A. Subsequent binding studies were performed employing the intrinsic tyrosine fluorescence properties of apoLp-III, which is highly quenched in the unbound state. A marked increase in tyrosine fluorescence intensity was observed upon binding to LPS or detoxified LPS, indicating a new microenvironment for Tyr-142. This also implies that the LPS carbohydrate region is involved in LPS binding. Dissociation constants (Kd) measured by apoLp-III titration were estimated at approximately 1 microM. Increasing the ionic strength did not decrease the Kd, neither did LPS phosphate removal. In addition, truncation apoLp-III mutants, lacking two complete helices, were still able to associate with LPS. This indicates that the association of apoLp-III with LPS may not be governed by charge but by hydrophobic interactions.

References

Aug 14, 1999·Biochimica Et Biophysica Acta·M NiereA Wiesner
Nov 6, 2003·Insect Biochemistry and Molecular Biology·Paul M M Weers, R O Ryan
Mar 23, 2006·Insect Biochemistry and Molecular Biology·Paul M M Weers, R O Ryan

Citations

Oct 21, 2010·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Agnieszka Zdybicka-Barabas, Małgorzata Cytryńska
Jul 13, 2014·Biochimica Et Biophysica Acta·Agnieszka Zdybicka-BarabasMałgorzata Cytryńska
Oct 16, 2015·Chemistry and Physics of Lipids·Jake ThistlePaul M M Weers
Aug 22, 2006·Biochemical and Biophysical Research Communications·Leonardo J LeonPaul M M Weers
Apr 21, 2016·Biochimica Et Biophysica Acta·Marta Palusińska-SzyszMałgorzata Cytryńska
Nov 22, 2011·Journal of Insect Physiology·Agnieszka Zdybicka-BarabasMałgorzata Cytryńska
Feb 20, 2013·Biochimica Et Biophysica Acta·Agnieszka Zdybicka-BarabasMałgorzata Cytryńska
Oct 29, 2014·Archives of Insect Biochemistry and Physiology·Qiu-Ning LiuChao-Liang Liu
Aug 23, 2020·International Journal of Molecular Sciences·Marta Palusińska-SzyszMałgorzata Cytryńska

Related Concepts

Apolipophorin-II
Metazoa
Apolipoproteins
SDS-PAGE
Fluorescence
Hydrogen-Ion Concentration
Kinetics
Lepidoptera
Lipopolysaccharides
Fluorescence Spectroscopy

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