Approaches to investigate the role of signaling in ER-to-Golgi transport

Methods Mol Biol. 2008:457:215-25. doi: 10.1007/978-1-59745-261-8_16.

Abstract

There have been many indications that kinases play a role in signaling the transport of proteins through the secretory pathway. Specifically, the serine/threonine kinase Akt affects the transport of cholesterol regulatory components from the endoplasmic reticulum (ER) to the Golgi. However, elucidating the target of Akt in this process has proven to be challenging. Here we describe an approach devised to investigate the Akt target(s) based on examining a potential candidate.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Biological Transport
  • Blotting, Western
  • CHO Cells
  • Conserved Sequence
  • Cricetinae
  • Cricetulus
  • Electrophoresis, Polyacrylamide Gel
  • Endoplasmic Reticulum / metabolism*
  • Evolution, Molecular
  • Golgi Apparatus / metabolism*
  • Humans
  • Immunoprecipitation
  • Microscopy
  • Molecular Biology / methods*
  • Phosphorylation
  • Proteins / chemistry
  • Proteins / metabolism
  • Proto-Oncogene Proteins c-akt / metabolism
  • Recombinant Fusion Proteins / metabolism
  • Sequence Alignment
  • Signal Transduction*

Substances

  • Proteins
  • Recombinant Fusion Proteins
  • Proto-Oncogene Proteins c-akt