Biochemistry of the Rap-specific guanine nucleotide exchange factors PDZ-GEF1 and -2

Methods Enzymol. 2006;407:174-86. doi: 10.1016/S0076-6879(05)07015-1.

Abstract

PDZ-GEFs represent one of four types of highly conserved Rap-specific guanine nucleotide exchange factors. They contain a number of well-known protein domains, including a "related to cyclic nucleotide binding domain" (RCBD), a PDZ-domain, a Ras-associating domain (RA), and, of course, a catalytic domain required for their exchange activity. Since their cloning more than 5 years ago, relatively little has been learned about their mode of regulation. Although their activity may in part depend on regulated membrane localization by means of the RA and/or PDZ domain, it seems highly likely that PDZ-GEFs can be modified by additional mechanisms as well. Based on analogy of the regulatory mechanisms of the cAMP-responsive GEF Epac, in the past we postulated a role for the RCBD domain in this. In this chapter, we give a detailed description of the methods that were used to unravel this mechanism in vitro and in vivo.

MeSH terms

  • Animals
  • Calorimetry
  • Cloning, Molecular
  • Escherichia coli / metabolism
  • Guanine Nucleotide Exchange Factors / analysis
  • Guanine Nucleotide Exchange Factors / metabolism*
  • Humans
  • Mice
  • NIH 3T3 Cells
  • Nerve Tissue Proteins / analysis
  • Nerve Tissue Proteins / metabolism*
  • Protein Structure, Tertiary
  • Rats
  • Spodoptera

Substances

  • Guanine Nucleotide Exchange Factors
  • Nerve Tissue Proteins
  • RAPGEF2 protein, human