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Title: Radioiodinated, photoactivatable phosphatidylcholine and phosphatidylserine: transfer properties and differential photoreactive interaction with human erythrocyte membrane proteins

Abstract

An isotopically labeled cross-linking reagent, succinimido 3-(3-(/sup 125/I)iodo-4-azidophenyl)propionate, has been synthesized and coupled to 1-acyl-2-(aminocaproyl)phosphatidylcholine according to previously described procedures. /sup 125/I- and N/sub 3/-labeled phosphatidylserine (/sup 125/I-N/sub 3/-PS) was produced from the phosphatidylcholine (PC) analog by phospholipase D catalyzed base exchange in the presence of L-serine. These phospholipid analogues are photoactivatable, are labeled with /sup 125/I at high specific activity, completely incorporate into synthetic vesicles, and spontaneously transfer between membranes. When an excess of acceptor vesicles or red blood cells (RBC) was mixed with a population of donor vesicles containing the /sup 125/I-N/sub 3/-phospholipids, approximately 40% of the analogues transferred to the acceptor population. After transfer in the dark to RBC, all of the /sup 125/I-N/sub 3/-PC incorporated into the cells could be removed by washing with serum, whereas the /sup 125/I-N/sub 3/-PS could not. After photolabeling of intact RBC, approx.50% of the PC and 20% of the PS cross-linked to membrane proteins as determined by their insolubility in CHCl/sub 3//MeOH. Analysis of probe distribution by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that /sup 125/I-N/sub 3/-PS preferentially labeled a M/sub r/ 30,000 peptide which contained approx.30% of the protein-bound label.

Authors:
; ;
Publication Date:
Research Org.:
Univ. of Texas M. D. Anderson Hospital and Tumor Institute, Houston
OSTI Identifier:
5942594
Resource Type:
Journal Article
Journal Name:
Biochemistry; (United States)
Additional Journal Information:
Journal Volume: 26:7
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; LABELLED COMPOUNDS; UPTAKE; PHOSPHOLIPIDS; AUTORADIOGRAPHY; CELL MEMBRANES; CHOLINE; ELECTROPHORESIS; ERYTHROCYTES; IODINE 125; ORGANIC IODINE COMPOUNDS; PEPTIDES; PHOTOCHEMISTRY; PROPIONIC ACID; SERINE; SODIUM IODIDES; ALCOHOLS; ALKALI METAL COMPOUNDS; AMINES; AMINO ACIDS; AMMONIUM COMPOUNDS; BETA DECAY RADIOISOTOPES; BIOLOGICAL MATERIALS; BLOOD; BLOOD CELLS; BODY FLUIDS; CARBOXYLIC ACIDS; CELL CONSTITUENTS; CHEMISTRY; DAYS LIVING RADIOISOTOPES; DRUGS; ELECTRON CAPTURE RADIOISOTOPES; ESTERS; HALIDES; HALOGEN COMPOUNDS; HYDROXY ACIDS; HYDROXY COMPOUNDS; INORGANIC PHOSPHORS; INTERMEDIATE MASS NUCLEI; IODIDES; IODINE COMPOUNDS; IODINE ISOTOPES; ISOTOPES; LIPIDS; LIPOTROPIC FACTORS; MATERIALS; MEMBRANES; MONOCARBOXYLIC ACIDS; NUCLEI; ODD-EVEN NUCLEI; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC HALOGEN COMPOUNDS; ORGANIC PHOSPHORUS COMPOUNDS; PHOSPHORS; PROTEINS; QUATERNARY COMPOUNDS; RADIOISOTOPES; SODIUM COMPOUNDS; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Schroit, A J, Madsen, J, and Ruoho, A E. Radioiodinated, photoactivatable phosphatidylcholine and phosphatidylserine: transfer properties and differential photoreactive interaction with human erythrocyte membrane proteins. United States: N. p., 1987. Web. doi:10.1021/bi00381a004.
Schroit, A J, Madsen, J, & Ruoho, A E. Radioiodinated, photoactivatable phosphatidylcholine and phosphatidylserine: transfer properties and differential photoreactive interaction with human erythrocyte membrane proteins. United States. https://doi.org/10.1021/bi00381a004
Schroit, A J, Madsen, J, and Ruoho, A E. 1987. "Radioiodinated, photoactivatable phosphatidylcholine and phosphatidylserine: transfer properties and differential photoreactive interaction with human erythrocyte membrane proteins". United States. https://doi.org/10.1021/bi00381a004.
@article{osti_5942594,
title = {Radioiodinated, photoactivatable phosphatidylcholine and phosphatidylserine: transfer properties and differential photoreactive interaction with human erythrocyte membrane proteins},
author = {Schroit, A J and Madsen, J and Ruoho, A E},
abstractNote = {An isotopically labeled cross-linking reagent, succinimido 3-(3-(/sup 125/I)iodo-4-azidophenyl)propionate, has been synthesized and coupled to 1-acyl-2-(aminocaproyl)phosphatidylcholine according to previously described procedures. /sup 125/I- and N/sub 3/-labeled phosphatidylserine (/sup 125/I-N/sub 3/-PS) was produced from the phosphatidylcholine (PC) analog by phospholipase D catalyzed base exchange in the presence of L-serine. These phospholipid analogues are photoactivatable, are labeled with /sup 125/I at high specific activity, completely incorporate into synthetic vesicles, and spontaneously transfer between membranes. When an excess of acceptor vesicles or red blood cells (RBC) was mixed with a population of donor vesicles containing the /sup 125/I-N/sub 3/-phospholipids, approximately 40% of the analogues transferred to the acceptor population. After transfer in the dark to RBC, all of the /sup 125/I-N/sub 3/-PC incorporated into the cells could be removed by washing with serum, whereas the /sup 125/I-N/sub 3/-PS could not. After photolabeling of intact RBC, approx.50% of the PC and 20% of the PS cross-linked to membrane proteins as determined by their insolubility in CHCl/sub 3//MeOH. Analysis of probe distribution by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that /sup 125/I-N/sub 3/-PS preferentially labeled a M/sub r/ 30,000 peptide which contained approx.30% of the protein-bound label.},
doi = {10.1021/bi00381a004},
url = {https://www.osti.gov/biblio/5942594}, journal = {Biochemistry; (United States)},
number = ,
volume = 26:7,
place = {United States},
year = {Tue Apr 07 00:00:00 EDT 1987},
month = {Tue Apr 07 00:00:00 EDT 1987}
}