New fluorescent probes reveal that flippase-mediated flip-flop of phosphatidylinositol across the endoplasmic reticulum membrane does not depend on the stereochemistry of the lipid. Academic Article uri icon

Overview

MeSH

  • 4-Chloro-7-nitrobenzofurazan
  • Animals
  • Dithionite
  • Magnetic Resonance Spectroscopy
  • Rats
  • Spectrometry, Mass, Electrospray Ionization
  • Stereoisomerism
  • Structure-Activity Relationship

MeSH Major

  • Endoplasmic Reticulum
  • Fluorescent Dyes
  • Phosphatidylinositols
  • Phospholipid Transfer Proteins

abstract

  • Glycerophospholipid flip-flop across biogenic membranes such as the endoplasmic reticulum (ER) is a fundamental feature of membrane biogenesis. Flip-flop requires the activity of specific membrane proteins called flippases. These proteins have yet to be identified in biogenic membranes and the molecular basis of their action is unknown. It is generally believed that flippase-facilitated glycerophospholipid flip-flop across the ER is governed by the stereochemistry of the glycerolipid, but this important issue has not been resolved. Here we investigate whether the ER flippase stereochemically recognizes the glycerophospholipids that it transports. To address this question we selected phosphatidylinositol (PI), a biologically important molecule with chiral centres in both its myo-inositol headgroup and its glycerol-lipid tail. The flip-flop of PI across the ER has not been previously reported. We synthesized fluorescence-labeled forms of all four diastereoisomers of PI and evaluated their flipping in rat liver ER vesicles, as well as in flippase-containing proteoliposomes reconstituted from a detergent extract of ER. Our results show that the flippase is able to translocate all four PI isomers and that both glycerol isomers of PI flip-flop across the ER membrane at rates similar to that measured for fluorescence-labeled phosphatidylcholine. Our data have important implications for recent hypotheses concerning the evolution of distinct homochiral glycerophospholipid membranes during the speciation of archaea and bacteria/eukarya from a common cellular ancestor.

publication date

  • April 7, 2005

has subject area

  • 4-Chloro-7-nitrobenzofurazan
  • Animals
  • Dithionite
  • Endoplasmic Reticulum
  • Fluorescent Dyes
  • Magnetic Resonance Spectroscopy
  • Phosphatidylinositols
  • Phospholipid Transfer Proteins
  • Rats
  • Spectrometry, Mass, Electrospray Ionization
  • Stereoisomerism
  • Structure-Activity Relationship

Research

keywords

  • Journal Article

Identity

Language

  • eng

Digital Object Identifier (DOI)

  • 10.1039/b500300h

PubMed ID

  • 15785818

Additional Document Info

start page

  • 1275

end page

  • 1283

volume

  • 3

number

  • 7