Interaction of leukotriene C4 and chinese hamster lung fibroblasts (V79A03 cells). 2. Subcellular distribution of binding and unlikely role of glutathione-S-transferase

Y. X. Liu, D. F. Contois, D. S. Watt, T. L. Walden, T. A. Fitz

Research output: Contribution to journalArticlepeer-review

Abstract

It was reported previously that radiation-induced cytotoxicity in V79A03 (V79) cells was attenuated by pretreatment of cells with leukotriene C4 (LTC4), leading us to determine that V79 cells possessed specific binding sites, with characteristics of receptors, for LTC4 (see the preceding, companion communication). Additional studies were conducted to determine the subcellular distribution and the chemical nature of the LTC4 binding site in V79 cells. Trypsin treatment of cells before LTC, binding assays resulted in a 74% reduction in high-affinity binding. In tests to examine the subcellular location of LTC4 binding, plasma membrane and nuclear fractions were obtained from V79 cells. In contrast to Scatchard analyses of LTC4 binding to intact cells which were curvilinear, Scatchard analyses of nuclear and plasma membrane fractions were linear, indicative of the presence in these cellular substituents of low and high-affinity binding, respectively. To examine the nature of the high-affinity LTC4 binding sites, intact V79 cells were photolyzed with [3H]-LTC4 rendered photoactive by preincubation with N-hydroxysuccinimidyl-4-azidobenzoate. The cell-bound radioactivity migrated during sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with an apparent molecular weight of approaximately 40 kdal. Five different commercial preparations of glutathione-S-transferase (GST), which has been implicated as a source of LTC4 "specific binding" in other cells, migrated in the same SDS_PAGE system with an apparent molecular weigth of 20-24 kdal. Furthermore, preincubations of V79 cells with three antisera generated against GST had minimal effects upon subsequent LTC4 binding to intact cells. These data, suggest that the radioprotective effect of LTC4 upon V79 cells may be attributable to a receptor-mediated phenomenon which appears distinct from leukotriene binding to GST.

Original languageEnglish
Pages (from-to)431-443
Number of pages13
JournalProstaglandins
Volume40
Issue number4
DOIs
StatePublished - Oct 1990

Bibliographical note

Funding Information:
The contributions of Dr. W.B. Jakoby for donation of antisera against GST, Mrs. Margaret Marr for expert cell culture, Ms. Yvonne Caicedo for technical manipulations, and Mrs. Jane Koeser for secretarial help, are gratefully acknowledged. This work was supported by NIH HD20780, USUHS Protocol C08517, and Defense Nuclear Agency Work Unit #B2152. The views presented in this paper are those of the authors. No endorsement by the Defense Nuclear Agency or the Department of Defense has been given or

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology

Fingerprint

Dive into the research topics of 'Interaction of leukotriene C4 and chinese hamster lung fibroblasts (V79A03 cells). 2. Subcellular distribution of binding and unlikely role of glutathione-S-transferase'. Together they form a unique fingerprint.

Cite this