Trp-21 is important in the processing and secretion of big endothelin-1

Adviye Ergul, David Puett

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

To investigate the intracellular processing of endothelin-1 (ET-1), the synthesis and secretion of preproET-1 (PPET-1) was studied in transiently transfected COS-7 cells. Replacements at the highly conserved C-terminal region of ET-1 were made by site-directed mutagenesis, with codons for residues Ile-20 and Trp-21 being replaced by one for Ala in the PPET-1 cDNA. The mutant and wildtype PPET-1 cDNAs were expressed in COS-7 cells following transient transfection, and cell media and extracts, collected after 48 h, were assayed for ET-1 and big ET-1 by radioimmunoassay. The concentration of immunoreactive ET-1 in the medium obtained from the Ala-21-ET-1 mutant was 10-fold lower than that from PPET-1 wildtype and (Ala-20-ET-1) PPET-1 transfected cells. Moreover, Ala-21 -big ET-1 accumulated in the cells transfected with the cDNA for (Ala-21-ET-1) PPET-1, whereas there was no significant accumulation of ET-1-like or big ET-1-like immunoreactivity in the cells transfected with cDNAs for PPET-1 wildtype and (Ala-20-ET-1) PPET-1. These results suggest that Trp-21 is involved in intracellular processing and secretion of big ET-1.

Original languageEnglish (US)
Pages (from-to)89-94
Number of pages6
JournalMolecular and Cellular Endocrinology
Volume110
Issue number1-2
DOIs
Publication statusPublished - Apr 28 1995

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Keywords

  • Big endothelin-1
  • Endothelin-1
  • Preproendothelin-1

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

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