Purification and functional properties of the membrane fissioning protein CtBP3/BARS

Methods Enzymol. 2005:404:296-316. doi: 10.1016/S0076-6879(05)04027-9.

Abstract

The fissioning protein CtBP3/BARS is a member of the CtBP transcription corepressor family of proteins. The characterization of this fissioning activity of CtBP3/BARS in both isolated Golgi membranes and in intact cells has indicated that the CtBP family includes multifunctional proteins that can act both in the nucleus and in the cytoplasm. The fissiogenic activity of CtBP3/BARS has a role in the fragmentation of the Golgi complex during mitosis and during intracellular membrane transport. This was demonstrated using a number of approaches and reagents, which are discussed in the following text, and which include recombinant proteins and mutants, antibodies, protein overexpression, RNA interference, antisense oligonucleotides, cell permeabilization, and electron miscroscopy, together with biochemical assays such as that for ADP-ribosylation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acyl Coenzyme A / metabolism
  • Acyltransferases / metabolism
  • Animals
  • Carrier Proteins / genetics
  • Carrier Proteins / immunology
  • Carrier Proteins / isolation & purification*
  • Carrier Proteins / physiology*
  • Golgi Apparatus / physiology
  • Humans
  • Mutation
  • NAD / metabolism
  • RNA, Small Interfering / genetics
  • Rats
  • Transcription Factors / genetics
  • Transcription Factors / immunology
  • Transcription Factors / isolation & purification*
  • Transcription Factors / physiology*
  • Transfection / methods

Substances

  • Acyl Coenzyme A
  • Carrier Proteins
  • Ctbp1 protein, rat
  • RNA, Small Interfering
  • Transcription Factors
  • NAD
  • Acyltransferases