Radioiodinated, photoactivatable phosphatidylcholine and phosphatidylserine: transfer properties and differential photoreactive interaction with human erythrocyte membrane proteins.
Radioiodinated, photoactivatable phosphatidylcholine and phosphatidylserine: transfer properties and differential photoreactive interaction with human erythrocyte membrane proteins.
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放射性碘化、光活化磷脂酰胆碱和磷脂酰丝氨酸：转移特性以及与人红细胞膜蛋白的差异光反应相互作用。
DOI：
10.1021/bi00381a004
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发表时间：
1987
期刊：
影响因子：
2.9
通讯作者：
Ruoho,AE
中科院分区：
文献类型：
--
作者：
Schroit,AJ;Madsen,J;Ruoho,AE
Department of Pharmacology, University of Wisconsin Medical School, Madison, Wisconsin 53701 Received September 10, 1986; Revised Manuscript Received November 7, 1986 abstract: An isotopically labeled cross-linking reagent, succinimido 3-(3-[125I] iodo-4-azidophenyl) propionate, has been synthesized and coupled to l-acyl-2-(aminocaproyl) phosphatidylcholine accordingto previously described procedures [Schroit, A. J., & Madsen, J.(1983) Biochemistry 22, 3617-3623]. 125I-and N3-labeled phosphatidylserine (125I-N3-PS) was produced from the phosphatidylcholine (PC) analogue by phospholipase D catalyzed base exchangein the presence of L-serine. These phospholipid analogues are photoactivatable, are labeled with 125I 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 125I-N3-phospholipids, approximately 40% of the analogues transferred to the acceptor population. After transfer in the dark to RBC, all of the 125I-N3-PC incorporated into the cells could be removed by washing with serum, whereas the 125I-N3-PS could not. After photolabeling of intact RBC,~ 50% of the PC and 20% of the PS cross-linked to membrane proteins as determined by their insolubility in CHCl3/MeOH. Analysis of probe distribution by sodiumdodecyl sulfate-polyacrylamide gel electrophoresis revealed that 125I-N3-PS preferentially labeled a Mr 30000 peptide which contained~ 30% of the protein-bound label.
