Assay and functional properties of PrBP(PDEdelta), a prenyl-binding protein interacting with multiple partners

Methods Enzymol. 2005;403:42-56. doi: 10.1016/S0076-6879(05)03005-3.

Abstract

A 17-kDa prenyl-binding protein, PrBP(PDEdelta), is highly conserved among various species from human to Caenorhabditis elegans. First identified as a putative regulatory delta subunit of the cyclic nucleotide phosphodiesterase (PDE6) purified from mammalian photoreceptor cells, PrBP(PDEdelta) has been hypothesized to reduce activation of PDE6 by the heterotrimeric G-protein, transducin, thereby desensitizing the photoresponse. However, recent work shows that PrBP(PDEdelta) interacts with numerous prenylated proteins at their farnesylated or geranylgeranylated C-termini, as well as with non-prenylated proteins. These polypeptides include small GTPases such as Rab13, Ras, Rap, and Rho6, as well as components involved in phototransduction (e.g., rod and cone PDE6, rod and cone opsin kinases). Expression of PrBP(PDEdelta) in tissues and organisms not expressing PDE6, the demonstration of multiple interacting partners with PrBP(PDEdelta), and its low abundance in rod outer segments all argue against it being a regulatory PDE6 subunit. This raises intriguing questions as to its physiological functions. In this chapter, we review the current status of PrBP(PDEdelta) and describe some of the assays used to determine these interactions in detail. In mammalian photoreceptors, the results are consistent with a role of PrBP(PDEdelta) in the transport of prenylated proteins from their site of synthesis in the inner segment to the outer segment where phototransduction occurs.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amphibians
  • Animals
  • Blotting, Western
  • Cattle
  • Cyclic Nucleotide Phosphodiesterases, Type 6
  • DNA, Complementary
  • Humans
  • Mice
  • Phosphoric Diester Hydrolases / metabolism*
  • Protein Binding
  • Reactive Oxygen Species
  • Retina / enzymology
  • Rod Cell Outer Segment / enzymology
  • Two-Hybrid System Techniques

Substances

  • DNA, Complementary
  • Reactive Oxygen Species
  • Phosphoric Diester Hydrolases
  • Cyclic Nucleotide Phosphodiesterases, Type 6
  • PDE6B protein, human
  • Pde6b protein, mouse