Measuring interactions of FERM domain-containing sorting nexin proteins with endosomal lipids and cargo molecules

Ghai, Rajesh, Mobli, Mehdi and Collins, Brett M. (2014) Measuring interactions of FERM domain-containing sorting nexin proteins with endosomal lipids and cargo molecules. Methods in Enzymology, 534 331-349. doi:10.1016/B978-0-12-397926-1.00019-6


Author Ghai, Rajesh
Mobli, Mehdi
Collins, Brett M.
Title Measuring interactions of FERM domain-containing sorting nexin proteins with endosomal lipids and cargo molecules
Journal name Methods in Enzymology   Check publisher's open access policy
ISSN 0076-6879
1557-7988
ISBN 9780123979261
Publication date 2014-01-01
Sub-type Article (original research)
DOI 10.1016/B978-0-12-397926-1.00019-6
Volume 534
Start page 331
End page 349
Total pages 19
Place of publication Maryland Heights, MO, United States
Publisher Academic Press
Language eng
Abstract Endosomal recycling pathways regulate cellular homeostasis via the transport of internalized material back to the plasma membrane. Phox homology (PX) and band 4.1/ezrin/radixin/moesin (FERM) domain-containing proteins are a recently identified subfamily of PX proteins that are critical for the recycling of numerous transmembrane cargo molecules. The PX-FERM subfamily includes three endosome-associated proteins called sorting nexin (SNX) 17, SNX27, and SNX31. These are modular peripheral membrane proteins that act as central scaffolds mediating protein-lipid interactions, cargo binding, and regulatory protein recruitment. This chapter outlines the methodology employed to classify the PX-FERM family using combined bioinformatics and structure prediction tools. It further details the application of isothermal titration calorimetry and nuclear magnetic resonance spectroscopy to understand the mechanisms that underpin their endosomal membrane recruitment and subsequent recognition of NPxY/NxxY peptide sorting motifs, present in many cargo receptors and required for their trafficking. It is now increasingly recognized that the formation of a stable trafficking complex is dictated by a multitude of coordinated protein-protein and protein-lipid interactions, and the approaches highlighted here will be useful for future studies aimed at understanding these biomolecular interactions in greater detail.
Keyword Band 4.1/ezrin/radixin/moesin domain
Endosome
Phosphoinositide
Phosphotyrosine-binding domain
Phox homology domain
Sorting nexin
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

 
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