Molecular mapping of pulmonary endothelial membrane glycoproteins of the intact rabbit lung. FASEB J 1990 Sep;4(12):3040-8
Date
09/01/1990Pubmed ID
2168327DOI
10.1096/fasebj.4.12.2168327Scopus ID
2-s2.0-0025204771 (requires institutional sign-in at Scopus site) 9 CitationsAbstract
To study the biochemical characteristics of endothelium in vivo, we radioiodinated endothelial membrane proteins of the perfused rabbit lung using a water soluble analog of the Bolton-Hunter reagent, 125I-sulfosuccinimidyl (hydroxyphenyl) propionate (125I-s-SHPP). This technique led to a 10-fold increase in specific activity of radioiodinated lung membrane protein compared with our previously reported method using lactoperoxidase and glucose oxidase-catalyzed radioiodination. Tissue autoradiography confirmed that radioiodination was largely confined to the endothelium. Perfusion pressure, wet-to-dry weight ratios, and the morphological appearance of the lungs were within normal limits, indicating that the procedure does not cause apparent lung injury. Lectin binding to a crude membrane fraction of 125I-s-SHPP labeled lung led to isolation of several putative endothelial membrane proteins. Immunoprecipitation studies with appropriate antibodies enabled the identification of radioiodinated angiotensin-converting enzyme and beta 2-microglobulin associated major histocompatibility complex class I molecules in the membrane fraction. This technique will be useful for studying biochemical responses of the endothelium in vivo to a variety of pharmacological and physiology stimuli.
Author List
Merker MP, Carley WW, Gillis CNMESH terms used to index this publication - Major topics in bold
AnimalsAutoradiography
Endothelium
Iodine Radioisotopes
Lung
Male
Membrane Glycoproteins
Peptidyl-Dipeptidase A
Precipitin Tests
Rabbits
Succinimides
