MURINE SPLENOCYTES EXPRESS A NALOXONE-INSENSITIVE BINDING-SITE FOR BETA-ENDORPHIN

被引:53
作者
SHAHABI, NA
LINNER, KM
SHARP, BM
机构
[1] HENNEPIN CTY MED CTR,DEPT MED,DIV ENDOCRINOL METAB & NUTR,701 PK AVE S,MINNEAPOLIS,MN 55415
[2] MINNEAPOLIS MED RES FDN INC,ENDOCRINE NEUROSCI RES LAB,MINNEAPOLIS,MN 55415
[3] UNIV MINNESOTA,MINNEAPOLIS,MN 55415
关键词
D O I
10.1210/endo-126-3-1442
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Naloxone-resistant binding sites for β-endor-phin have previously been observed on transformed peripheral blood mononuclear cells and on the EL4-thymoma cell line. These sites may be related to the naloxone-insensitive immunomodulatory effects of β-endorphin. The present study was performed 1) to determine whether these sites are present on normal splenocytes and 2) to characterize them. Ficoll-hypaque-purified murine splenocytes were used in a RRA with [125I] β-endorphin. Neither fresh intact cells obtained from viral antibody-free mice nor membrane preparations showed evidence of binding. How-ever, splenocytes cultured in 5% fetal bovine serum for 24-96 h showed sites on intact cells or membranes (after 3 h in culture no sites were present). Intact cultured splenocytes demonstrated a saturable binding isotherm, and Scatchard analysis showed a single site (Kd= 4.1 X 10-9 M). Competition studies showed that N-acetyl-β-endorphin (N-Ac-β-endorphin)-(1-31) was equipotent to ?-endorphin-(1-31). β-Endorphin-(6-31) and β-endor-phin-(28-31) were approximately 10- and 1000-fold less potent, respectively, whereas β-endorphin-(1-27) and naloxone were completely ineffective. Covalent cross-linking of [125I] β-endor-phin to splenocytes and resolution by gel electrophoresis showed bands at 66K and 57K which were displaced equipotently by increasing amounts of (8-endorphin and N-Ac-β-endorphin. β-Endorphin-(18-31) or (28-31) were less potent, and naloxone or other opioid ligands selective for receptor subtypes were ineffective. Thus, high affinity, naloxone-insensitive binding sites for (β-endorphin, which show competition characteristics distinctively different from brain opiate receptors, are inducible on normal mouse splenocytes. N-Ac-β-endorphin, presumed to be an inactivation product of β-endorphin because it fails to bind brain opiate receptors, may be functional at this naloxone-insensitive binding site. © 1990 by The Endocrine Society.
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页码:1442 / 1448
页数:7
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