A fluorescent glycolipid-binding peptide probe traces cholesterol dependent microdomain-derived trafficking pathways.

PloS One
Steffen SteinertRachel Kraut

Abstract

The uptake and intracellular trafficking of sphingolipids, which self-associate into plasma membrane microdomains, is associated with many pathological conditions, including viral and toxin infection, lipid storage disease, and neurodegenerative disease. However, the means available to label the trafficking pathways of sphingolipids in live cells are extremely limited. In order to address this problem, we have developed an exogenous, non-toxic probe consisting of a 25-amino acid sphingolipid binding domain, the SBD, derived from the amyloid peptide Abeta, and conjugated by a neutral linker with an organic fluorophore. The current work presents the characterization of the sphingolipid binding and live cell trafficking of this novel probe, the SBD peptide. SBD was the name given to a motif originally recognized by Fantini et al in a number of glycolipid-associated proteins, and was proposed to interact with sphingolipids in membrane microdomains. In accordance with Fantini's model, optimal SBD binding to membranes depends on the presence of sphingolipids and cholesterol. In synthetic membrane binding assays, SBD interacts preferentially with raft-like lipid mixtures containing sphingomyelin, cholesterol, and complex gangliosides ...Continue Reading

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Citations

Aug 20, 2014·BioMed Research International·Ravi Kant Upadhyay
Apr 21, 2009·PMC Biophysics·Guillaume Tresset
Feb 20, 2010·BMC Cell Biology·Matthew W ConklinPatricia J Keely
Jan 11, 2011·Journal of Neurochemistry·Rachel Kraut
Jan 11, 2011·Journal of Neurochemistry·Maria Dolores LedesmaEdward H Schuchman
Dec 18, 2009·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Feb 27, 2019·Journal of Industrial Microbiology & Biotechnology·Miguel C Santoscoy, Laura R Jarboe
Mar 25, 2015·Biological Chemistry·Johannes KornhuberChristiane Mühle
Jan 10, 2018·Chemical & Pharmaceutical Bulletin·Kaori Sakurai
Jun 20, 2013·Chembiochem : a European Journal of Chemical Biology·Bridget L Stocker, Mattie S M Timmer
Jul 7, 2018·Chemical Reviews·Carmanah D HunterChristopher W Cairo
Jul 6, 2010·Matrix Biology : Journal of the International Society for Matrix Biology·Henry J GreynerMark E Mummert

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Datasets Mentioned

BETA
GM95

Methods Mentioned

BETA
fluorescence imaging
Fluorescence
surface plasmon resonance
transfection
GTPases
enzymatic assay
atomic force microscopy
FCS
dot blots
chip

Software Mentioned

Wavemetrics
Adobe Photoshop
Origin
Quantity One
Igor Pro
ImageJ
Systat
SigmaPlot

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