Radiolabeling of the hydrophobic components of lung surfactant with 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine

Details

Serval ID
serval:BIB_2C44AB587FEE
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Radiolabeling of the hydrophobic components of lung surfactant with 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine
Journal
Analytical Biochemistry
Author(s)
Gilliard  N., Pappert  D., Merritt  T. A., Heldt  G., Spragg  R. G.
ISSN
0003-2697 (Print)
Publication state
Published
Issued date
03/1991
Volume
193
Number
2
Pages
310-5
Notes
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S. --- Old month value: Mar 2
Abstract
Studies of the metabolism and distribution of lung surfactant are aided by use of radiolabeled surfactant or surfactant components. These studies have often made use of [3H]- or [14C]phosphatidylcholine. Analysis of the lung content of surfactant containing these beta-emitting labels usually requires tissue digestion, use of scintillation fluids, and significant correction for quenching of photon production. Because use of a gamma-emitting isotope would obviate these requirements, we have investigated the use of 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine ([125I]TID), a lipophilic photoactivatable compound, to radiolabel pulmonary surfactant. Our results indicate that, during photoactivation, products of [125I]TID are produced that result in radiolabeling of both the lipid and protein components of extracted porcine surfactant. Separation of radiolabeled surfactant from hydrophobic nonlabelling photolysis products was accomplished by gel chromatography. Exposure of surfactant (34 mumol/ml) to [125I]TID under labeling conditions resulted in incorporation of 45.3 +/- 5.1% of the radiolabel. Incorporation of radiolabel in the various phospholipids of lung surfactant was approximately equivalent. Lipophilic surfactant apoproteins were also radiolabeled. Finally, both in vitro and in vivo testing of radiolabeled surfactant (0.1 microCi/mg) revealed full retention of surface tension lowering ability.
Keywords
Animals Apoproteins/analysis Azirines/*diagnostic use Chromatography, Gel Chromatography, Thin Layer Electrophoresis, Polyacrylamide Gel Iodine Radioisotopes/*diagnostic use Isotope Labeling/methods Light Phospholipids/analysis *Pulmonary Surfactants/chemistry Swine
Pubmed
Web of science
Create date
28/01/2008 10:50
Last modification date
20/08/2019 13:11
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