Raft Association of SNAP Receptors Acting in Apical Trafficking in Madin-Darby Canine Kidney Cells
Overview
Affiliations
We have investigated the relationships between the apical sorting mechanism using lipid rafts and the soluble N-ethyl maleimide-sensitive factor attachment protein receptor (SNARE) machinery, which is involved in membrane docking and fusion. We first confirmed that anti-alpha-SNAP antibodies inhibit the apical pathway in Madin- Darby canine kidney (MDCK) cells; in addition, we report that a recombinant SNAP protein stimulates the apical transport whereas a SNAP mutant inhibits this transport step. Based on t-SNARE overexpression experiments and the effect of botulinum neurotoxin E, syntaxin 3 and SNAP-23 have been implicated in apical membrane trafficking. Here, we show in permeabilized MDCK cells that antisyntaxin 3 and anti-SNAP-23 antibodies lower surface delivery of an apical reporter protein. Moreover, using a similar approach, we show that tetanus toxin-insensitive, vesicle-associated membrane protein (TI-VAMP; also called VAMP7), a recently described apical v-SNARE, is involved. Furthermore, we show the presence of syntaxin 3 and TI-VAMP in isolated apical carriers. Polarized apical sorting has been postulated to be mediated by the clustering of apical proteins into dynamic sphingolipid-cholesterol rafts. We provide evidence that syntaxin 3 and TI-VAMP are raft-associated. These data support a raft-based mechanism for the sorting of not only apically destined cargo but also of SNAREs having functions in apical membrane-docking and fusion events.
Partitioning to ordered membrane domains regulates the kinetics of secretory traffic.
Castello-Serrano I, Heberle F, Diaz-Rohrer B, Ippolito R, Shurer C, Lujan P Elife. 2024; 12.
PMID: 38837189 PMC: 11152573. DOI: 10.7554/eLife.89306.
Membrane lipid rafts are required for AMPA receptor tyrosine phosphorylation.
Hayashi T Front Synaptic Neurosci. 2022; 14:921772.
PMID: 36387774 PMC: 9662747. DOI: 10.3389/fnsyn.2022.921772.
Ye W, Zhu D, Xu B, Xu M, Liu Y, Xie J Transl Cancer Res. 2022; 8(2):466-472.
PMID: 35116778 PMC: 8798739. DOI: 10.21037/tcr.2019.02.11.
Kiyoshi C, Tedeschi A Dev Neurobiol. 2020; 80(7-8):277-301.
PMID: 32902152 PMC: 7754183. DOI: 10.1002/dneu.22780.
The ins and outs of the Arf4-based ciliary membrane-targeting complex.
Deretic D, Lorentzen E, Fresquez T Small GTPases. 2019; 12(1):1-12.
PMID: 31068062 PMC: 7781591. DOI: 10.1080/21541248.2019.1616355.