Supported lipid bilayer array to study clathrin mediated endocytosis in vitro

Hiroaki Suzuki, Thomas Pucadyil, Rajesh Ramachandran, Shoji Takeuchi, Sandra L. Schmid

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

We describe our effort to realize an assay system to study clathrin-mediated endocytosis (CME) in vitro. Since CME related proteins are known to bind phosphatidylinositol-4,5-bisphosphate (PI(4,5)P2), we propose an accessible method to create an array of supported lipid bilayers (SLBs) with different lipid compositions, using a PDMS stamp and a microchannel. Formation of a uniform SLB of a complex lipid mixture containing PI(4,5)P 2 was confirmed by fluorescent imaging and fluorescence recovery after photobleaching (FRAP). Our technique should allow us to vary the concentration of PI(4,5)P2 or other membrane constituents to study their requirements. In preliminary experiments we showed that dynamin binds to the planar bilayer containing PI(4,5)P2, but without causing membrane tubulation as it does when dynamin binds liposomes.

Original languageEnglish (US)
Title of host publicationProceedings of the IEEE International Conference on Micro Electro Mechanical Systems (MEMS)
Pages481-484
Number of pages4
StatePublished - 2007
Event20th IEEE International Conference on Micro Electro Mechanical Systems, MEMS 2007 - Kobe, Japan
Duration: Jan 21 2007Jan 25 2007

Other

Other20th IEEE International Conference on Micro Electro Mechanical Systems, MEMS 2007
CountryJapan
CityKobe
Period1/21/071/25/07

Fingerprint

Dynamins
Clathrin
Lipid bilayers
Lipids
lipids
Membranes
Photobleaching
Liposomes
Phosphatidylinositols
Microchannels
Assays
Fluorescence
membranes
Proteins
Imaging techniques
Recovery
microchannels
Chemical analysis
Experiments
recovery

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Mechanical Engineering
  • Condensed Matter Physics
  • Electronic, Optical and Magnetic Materials

Cite this

Suzuki, H., Pucadyil, T., Ramachandran, R., Takeuchi, S., & Schmid, S. L. (2007). Supported lipid bilayer array to study clathrin mediated endocytosis in vitro. In Proceedings of the IEEE International Conference on Micro Electro Mechanical Systems (MEMS) (pp. 481-484). [4433076]

Supported lipid bilayer array to study clathrin mediated endocytosis in vitro. / Suzuki, Hiroaki; Pucadyil, Thomas; Ramachandran, Rajesh; Takeuchi, Shoji; Schmid, Sandra L.

Proceedings of the IEEE International Conference on Micro Electro Mechanical Systems (MEMS). 2007. p. 481-484 4433076.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Suzuki, H, Pucadyil, T, Ramachandran, R, Takeuchi, S & Schmid, SL 2007, Supported lipid bilayer array to study clathrin mediated endocytosis in vitro. in Proceedings of the IEEE International Conference on Micro Electro Mechanical Systems (MEMS)., 4433076, pp. 481-484, 20th IEEE International Conference on Micro Electro Mechanical Systems, MEMS 2007, Kobe, Japan, 1/21/07.
Suzuki H, Pucadyil T, Ramachandran R, Takeuchi S, Schmid SL. Supported lipid bilayer array to study clathrin mediated endocytosis in vitro. In Proceedings of the IEEE International Conference on Micro Electro Mechanical Systems (MEMS). 2007. p. 481-484. 4433076
Suzuki, Hiroaki ; Pucadyil, Thomas ; Ramachandran, Rajesh ; Takeuchi, Shoji ; Schmid, Sandra L. / Supported lipid bilayer array to study clathrin mediated endocytosis in vitro. Proceedings of the IEEE International Conference on Micro Electro Mechanical Systems (MEMS). 2007. pp. 481-484
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