Adaptor and clathrin exchange at the plasma membrane and trans-Golgi network

Mol Biol Cell. 2003 Feb;14(2):516-28. doi: 10.1091/mbc.e02-06-0353.

Abstract

We previously demonstrated, using fluorescence recovery after photobleaching, that clathrin in clathrin-coated pits at the plasma membrane exchanges with free clathrin in the cytosol, suggesting that clathrin-coated pits are dynamic structures. We now investigated whether clathrin at the trans-Golgi network as well as the clathrin adaptors AP2 and AP1 in clathrin-coated pits at the plasma membrane and trans-Golgi network, respectively, also exchange with free proteins in the cytosol. We found that when the budding of clathrin-coated vesicle is blocked without significantly affecting the structure of clathrin-coated pits, both clathrin and AP2 at the plasma membrane and clathrin and AP1 at the trans-Golgi network exchange rapidly with free proteins in the cytosol. In contrast, when budding of clathrin-coated vesicles was blocked at the plasma membrane or trans-Golgi network by hypertonic sucrose or K(+) depletion, conditions that markedly affect the structure of clathrin-coated pits, clathrin exchange was blocked but AP2 at the plasma membrane and both AP1 and the GGA1 adaptor at the trans-Golgi network continue to rapidly exchange. We conclude that clathrin-coated pits are dynamic structures with rapid exchange of both clathrin and adaptors and that adaptors are able to exchange independently of clathrin when clathrin exchange is blocked.

MeSH terms

  • Adaptor Protein Complex 1 / metabolism
  • Adaptor Protein Complex 2 / metabolism
  • Cell Membrane / metabolism*
  • Clathrin / metabolism*
  • Cytosol / metabolism
  • DNA / metabolism
  • Green Fluorescent Proteins
  • HeLa Cells
  • Humans
  • Kinetics
  • Luminescent Proteins / metabolism
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Potassium / metabolism
  • Temperature
  • Time Factors
  • Transfection
  • trans-Golgi Network / metabolism*

Substances

  • Adaptor Protein Complex 1
  • Adaptor Protein Complex 2
  • Clathrin
  • Luminescent Proteins
  • Green Fluorescent Proteins
  • DNA
  • Potassium