Two mechanistically distinct forms of endocytosis in adrenal chromaffin cells: Differential effects of SH3 domains and amphiphysin antagonism

FEBS Lett. 2006 May 29;580(13):3263-9. doi: 10.1016/j.febslet.2006.04.083. Epub 2006 May 4.

Abstract

We previously identified two forms of endocytosis using capacitance measurements in chromaffin cells: rapid endocytosis (RE), dynamin-1 dependent but clathrin-independent and slow endocytosis (SE), dynamin-2 and clathrin-dependent. Various recombinant SH3 domains that interact with the proline-rich domain of dynamin were introduced into single cells via the patch pipette. GST-SH3 domains of amphiphysin-1, intersectin-IC, and endophilin-I inhibited SE but had no effect on RE. Grb2-SH3 (N-terminal) or a mutant of amphiphysin-1-SH3 was inactive on either process. These data confirm that dynamin-1 dependent RE is independent of clathrin and show that amphiphysin is exclusively associated with clathrin and dynamin-2-dependent SE.

Publication types

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

MeSH terms

  • Acyltransferases / genetics
  • Acyltransferases / metabolism
  • Adaptor Proteins, Vesicular Transport / genetics
  • Adaptor Proteins, Vesicular Transport / metabolism
  • Adrenal Glands / cytology*
  • Adrenal Glands / physiology
  • Animals
  • Antibodies / pharmacology
  • Cattle
  • Chromaffin Cells / drug effects
  • Chromaffin Cells / metabolism
  • Chromaffin Cells / physiology*
  • Clathrin / metabolism
  • Dynamin I / metabolism
  • Dynamin II / metabolism
  • Electric Capacitance
  • Endocytosis / drug effects
  • Endocytosis / physiology*
  • Nerve Tissue Proteins / antagonists & inhibitors
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism*
  • Recombinant Proteins / pharmacology
  • Surface Plasmon Resonance
  • src Homology Domains / genetics
  • src Homology Domains / physiology*

Substances

  • Adaptor Proteins, Vesicular Transport
  • Antibodies
  • Clathrin
  • Nerve Tissue Proteins
  • Recombinant Proteins
  • intersectin 1
  • amphiphysin
  • Acyltransferases
  • 2-acylglycerophosphate acyltransferase
  • Dynamin I
  • Dynamin II