A novel method to isolate and map endothelial membrane proteins from pulmonary vasculature. Academic Article uri icon

Overview

MeSH

  • Animals
  • Antibodies, Monoclonal
  • Chromatography, Affinity
  • Female
  • Immunoblotting
  • Immunohistochemistry
  • Rats
  • Rats, Sprague-Dawley

MeSH Major

  • Biotinylation
  • Cell Membrane
  • Endothelium, Vascular
  • Lung
  • Membrane Proteins

abstract

  • Vascular endothelium has attracted extensive attention due to its important role in many physiological and pathological processes. Many methods have been developed to study the components and their functions in vascular endothelium. Here we report a novel approach to investigate vascular endothelium using normal rat lungs as the model. We perfused lung vascular beds with sulfosuccinimidyl-6-(biotinamido) hexanoate, a biotin analog, to label endothelial membrane proteins. The biotinylated proteins were isolated from lung homogenate with immobilized monomeric avidin and confirmed to be highly pure endothelial membrane proteins with little contamination of intracellular proteins. These biotinylated proteins were used as immunogens for development of monoclonal antibodies. Indeed, newly generated monoclonal antibodies have revealed different expression patterns of proteins across tissues. Some proteins were found highly specifically expressed to capillary vessels of pulmonary vasculature. This method has also been proven useful for investigating vasculature of other organs, as this study explored.

publication date

  • April 2005

has subject area

  • Animals
  • Antibodies, Monoclonal
  • Biotinylation
  • Cell Membrane
  • Chromatography, Affinity
  • Endothelium, Vascular
  • Female
  • Immunoblotting
  • Immunohistochemistry
  • Lung
  • Membrane Proteins
  • Rats
  • Rats, Sprague-Dawley

Research

keywords

  • Journal Article

Identity

Language

  • eng

Digital Object Identifier (DOI)

  • 10.1152/ajpcell.00262.2004

PubMed ID

  • 15590900

Additional Document Info

start page

  • C950

end page

  • C956

volume

  • 288

number

  • 4