NDLI: A new approach to the molecular analysis of docking, priming, and regulated membrane fusion

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Hill coefficients of dietary polyphenolic enzyme inhibitiors: can beneficial health effects of dietary polyphenols be explained by allosteric enzyme denaturing?

A new approach to the molecular analysis of docking, priming, and regulated membrane fusion

JOCB Bulletin ( Journal of Chemical Biology , Volume 4 , Issue 3 )

Journal of Chemical Biology : Volume 4, Issue 2, April 2011

Journal of Chemical Biology : Volume 4, Issue 1, January 2011

Journal of Chemical Biology : Volume 3

Journal of Chemical Biology : Volume 2

Journal of Chemical Biology : Volume 1

A new approach to the molecular analysis of docking, priming, and regulated membrane fusion

Content Provider	Springer Nature Link
Author	Rogasevskaia, Tatiana P. Coorssen, Jens R.
Copyright Year	2011
Abstract	Studies using isolated sea urchin cortical vesicles have proven invaluable in dissecting mechanisms of Ca$^{2+}$-triggered membrane fusion. However, only acute molecular manipulations are possible in vitro. Here, using selective pharmacological manipulations of sea urchin eggs ex vivo, we test the hypothesis that specific lipidic components of the membrane matrix selectively affect defined late stages of exocytosis, particularly the Ca$^{2+}$-triggered steps of fast membrane fusion. Egg treatments with cholesterol-lowering drugs resulted in the inhibition of vesicle fusion. Exogenous cholesterol recovered fusion extent and efficiency in cholesterol-depleted membranes; α-tocopherol, a structurally dissimilar curvature analogue, selectively restored fusion extent. Inhibition of phospholipase C reduced vesicle phosphatidylethanolamine and suppressed both the extent and kinetics of fusion. Although phosphatidylinositol-3-kinase inhibition altered levels of polyphosphoinositide species and reduced all fusion parameters, sequestering polyphosphoinositides selectively inhibited fusion kinetics. Thus, cholesterol and phosphatidylethanolamine play direct roles in the fusion pathway, contributing negative curvature. Cholesterol also organizes the physiological fusion site, defining fusion efficiency. A selective influence of phosphatidylethanolamine on fusion kinetics sheds light on the local microdomain structure at the site of docking/fusion. Polyphosphoinositides have modulatory upstream roles in priming: alterations in specific polyphosphoinositides likely represent the terminal priming steps defining fully docked, release-ready vesicles. Thus, this pharmacological approach has the potential to be a robust high-throughput platform to identify molecular components of the physiological fusion machine critical to docking, priming, and triggered fusion.
Starting Page	117
Ending Page	136
Page Count	20
File Format	PDF
ISSN	18646158
Journal	Journal of Chemical Biology
Volume Number	4
Issue Number	3
e-ISSN	18646166
Language	English
Publisher	Springer-Verlag
Publisher Date	2011-02-08
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	Sea urchin eggs Cortical vesicles Ca$^{2+}$-triggered fusion Priming Cholesterol Phosphatidylethanolamine Polyphosphoinositides Biophysics and Biological Physics Cell Biology Physical Chemistry Biochemistry Pharmacology/Toxicology
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Biophysics

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