Single-molecule analysis of a molecular disassemblase reveals the mechanism of Hsc70-driven clathrin uncoating

Nat Struct Mol Biol. 2011 Mar;18(3):295-301. doi: 10.1038/nsmb.1985. Epub 2011 Jan 30.

Abstract

Heat shock cognate protein-70 (Hsc70) supports remodeling of protein complexes, such as disassembly of clathrin coats on endocytic coated vesicles. To understand how a simple ATP-driven molecular clamp catalyzes a large-scale disassembly reaction, we have used single-particle fluorescence imaging to track the dynamics of Hsc70 and its clathrin substrate in real time. Hsc70 accumulates to a critical level, determined by kinetic modeling to be one Hsc70 for every two functional attachment sites; rapid, all-or-none uncoating then ensues. We propose that Hsc70 traps conformational distortions, seen previously by cryo-EM, in the vicinity of each occupied site and that accumulation of local strains destabilizes the clathrin lattice. Capture of conformational fluctuations may be a general mechanism for chaperone-driven disassembly of protein complexes.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Animals
  • Auxilins / metabolism
  • Binding Sites
  • Cattle
  • Clathrin / chemistry
  • Clathrin / genetics
  • Clathrin / metabolism*
  • Clathrin-Coated Vesicles / chemistry
  • Clathrin-Coated Vesicles / genetics
  • Clathrin-Coated Vesicles / metabolism*
  • Escherichia coli / genetics
  • Fluorescence
  • Gene Expression
  • HSC70 Heat-Shock Proteins / chemistry
  • HSC70 Heat-Shock Proteins / genetics
  • HSC70 Heat-Shock Proteins / metabolism*
  • Insecta / cytology
  • Kinetics
  • Models, Molecular
  • Mutation
  • Rats

Substances

  • Auxilins
  • Clathrin
  • HSC70 Heat-Shock Proteins
  • Adenosine Triphosphate