DOI: 10.1073/pnas.96.7.3734. Raft association of SNAP receptors acting in apical trafficking in Madin

Raft association of SNAP receptors acting in apical trafficking in Madin–Darby canine kidney cells

10.1073/pnas.96.7.3734

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

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.

Bibliography

Lafont, F., Verkade, P., Galli, T., Wimmer, C., Louvard, D., & Simons, K. (1999). Raft association of SNAP receptors acting in apical trafficking in MadinâDarby canine kidney cells. Proceedings of the National Academy of Sciences, 96(7), 3734â3738.

Authors 6

Frank Lafont (first)
Paul Verkade (additional)
Thierry Galli (additional)
Christian Wimmer (additional)
Daniel Louvard (additional)
Kai Simons (additional)

References 39 Referenced 198

10.1146/annurev.cb.08.110192.002143
10.1016/S0962-8924(97)01130-6
P Scheiffele, K Simons Epithelial Morphogenesis in Development and Disease, eds W Birchmeier, C Birchmeier (Harwood, New York, , in press. (1998). / Epithelial Morphogenesis in Development and Disease by Scheiffele P (1998)
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F Lafont, E Ikonen, K Simons Cell Biology: A Laboratory Handbook, ed J E Celis (Academic, San Diego) 2, 227–236 (1998). / Cell Biology: A Laboratory Handbook by Lafont F (1998)
10.1016/0003-2697(84)90782-6
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10.1242/jcs.111.17.2681

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:39 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 5:30 p.m.)
Indexed	3 weeks, 4 days ago (Aug. 7, 2025, 4:51 a.m.)
Issued	26 years, 5 months ago (March 30, 1999)
Published	26 years, 5 months ago (March 30, 1999)
Published Online	26 years, 5 months ago (March 30, 1999)
Published Print	26 years, 5 months ago (March 30, 1999)

Funders 0

None

BibTeX

@article{Lafont_1999, title={Raft association of SNAP receptors acting in apical trafficking in Madin–Darby canine kidney cells}, volume={96}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.96.7.3734}, DOI={10.1073/pnas.96.7.3734}, number={7}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Lafont, Frank and Verkade, Paul and Galli, Thierry and Wimmer, Christian and Louvard, Daniel and Simons, Kai}, year={1999}, month=mar, pages={3734–3738} }

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