Testing Pollen Tube Proteins for In Vivo Binding to Phosphatidic Acid by n-Butanol Treatment and Confocal Microscopy.

Methods in Molecular Biology
Carolin Fritz, Benedikt Kost

Abstract

The general role of cellular membranes is to provide a barrier and to generate separate reaction spaces. However, additional functions of membrane domains enriched in certain classes of lipids have been discovered, which represent an important area of ongoing research. Such membrane domains can be found in cells at different size scales (e.g., nanodomains, microdomains), represent membrane regions with special physical properties and play important roles in the direct or indirect propagation of signaling processes. Domain formation within the plasma membrane (PM) does not only involve the accumulation of specific lipids, but also the recruitment of specific transmembrane or PM-associated peripheral proteins. Phosphatidic acid (PA) is increasingly recognized as an important signaling lipid and component of PM domains. This lipid is involved in the regulation not only of biotic or abiotic stress responses, but also of pollen tube tip growth and of other forms of polar cell expansion. Although many PA-binding proteins have been characterized, a conserved PA interaction motif could not be identified in these proteins. Consequently, protein binding to PA cannot be predicted based on sequence analysis, but has to be biochemically tes...Continue Reading

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