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In vivo and in vitro Studies of Adaptor-clathrin Interaction
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In vivo and in vitro Studies of Adaptor-clathrin Interaction

In vivo and in vitro Studies of Adaptor-clathrin Interaction

DOI:
10.3791/2352-v

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17:14 min

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January 26, 2011

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, , ,
1Department of Biochemistry and Molecular Biology,Colorado State University

Chapters

  • 01:34Introduction
  • 02:10Generation of GFP-tagged Sla1 and Sla1 Clatherin-binding Mutant Yeast Strains
  • 03:22Fluorescence Microscopy
  • 05:09Fluorescence Image Analysis
  • 06:41Preparation of Yeast Cytosolic Extract
  • 09:54GST-fusion Protein Affinity Assay
  • 11:53Preparation of Total Cell Extract
  • 13:36Co-immunoprecipitation Assay
  • 15:16Representative Results
  • 16:39Conclusion

Summary

Automatic Translation

Automatic Translation

Clathrin-mediated endocytosis depends on adaptor proteins that coordinate cargo selection and clathrin coat assembly. Here we describe procedures to study adaptor-clathrin physical interaction and live cell imaging approaches using as a model the yeast endocytic adaptor protein Sla1p.

Tags

In VivoIn VitroAdaptor-clathrin InteractionEndocytic PathwayClathrin-coated Vesicles (CCVs)Clathrin LatticeClathrin TriskeliaClathrin-binding AdaptorsMembrane ProteinsTransmembrane Protein CargoCCV Formation MachineryClathrin AdaptorsN-terminal -propeller DomainModular ProteinsLinker Regions

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