Direct role of furin in mammalian prosomatostatin processing

Aristea S. Galanopoulou, N. G. Seidah, Y. C. Patel

Research output: Contribution to journalArticle

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Abstract

We have previously reported that rat prosomatostatin (rPSS) undergoes conversion at Arg(↓) and Lys(↓) monobasic sites to SS-28 and PSS-(1-10) respectively in COS-7 cells, and have proposed furin or a related enzyme of the constitutive secretory pathway as the endoproteinase responsible. Here we have tested directly the ability of furin to cleave rPSS at the two monobasic sites as well as at the RXRK dibasic site of SS-14 conversion (a furin motif, except for Lys substituting for Arg at P1). Recombinant vaccinia virus (VV) vectors were used to co-express rPSS with graded doses of furin in COS-7 cells and LoVo colon carcinoma cells deficient in furin. PSS and cleavage products in cell extracts and media were characterized by HPLC analysis and C-terminal [SS-14-like immunoreactivity (SS-14 LI)] and N-terminal [PSS-(1-10) LT] directed radioimmunoassays. There was a dose-dependent increase in SS-28 production from rPSS by furin in COS-7 cells from 29% (control) to 58% (high-dose furin) associated with a progressive decrease in unprocessed PSS from > 60% to ~ 20% of total SS-14 LI. Significant SS-14 production occurred only at high levels of furin infection. Control LoVo cells infected with VV:rPSS exhibited production of ~ 21% SS-28, ~ 15% PSS-(1-10) and 3.5% SS-14. Infection of LoVo cells with VV:hfurin (hfurin = human furin) enhanced SS-28 production to 30-34%. SS-14 synthesis also increased to 25-40%, probably by conversion from SS-28. Overexpression of furin in COS-7 or LoVo cells failed to increase PSS-(1-10) production. These results show that furin is a candidate SS-28 convertase. Arginine is the preferred residue at the P1 site of furin cleavage. Furin does not process rPSS to PSS-(1-10), suggesting the existence of another monobasic convertase with a preference for Lys rather than Arg at P1. Such an enzyme could also explain the presence of endogenous SS-28-, PSS-(1-10)- and SS-14-producing activities in LoVo cells.

Original languageEnglish (US)
Pages (from-to)33-40
Number of pages8
JournalBiochemical Journal
Volume309
Issue number1
StatePublished - 1995
Externally publishedYes

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Furin
Processing
Rats
Vaccinia virus
COS Cells
Viruses
prosomatostatin
Cells
Secretory Pathway
Enzymes
Infection
Cell Extracts

ASJC Scopus subject areas

  • Biochemistry

Cite this

Direct role of furin in mammalian prosomatostatin processing. / Galanopoulou, Aristea S.; Seidah, N. G.; Patel, Y. C.

In: Biochemical Journal, Vol. 309, No. 1, 1995, p. 33-40.

Research output: Contribution to journalArticle

Galanopoulou, AS, Seidah, NG & Patel, YC 1995, 'Direct role of furin in mammalian prosomatostatin processing', Biochemical Journal, vol. 309, no. 1, pp. 33-40.
Galanopoulou, Aristea S. ; Seidah, N. G. ; Patel, Y. C. / Direct role of furin in mammalian prosomatostatin processing. In: Biochemical Journal. 1995 ; Vol. 309, No. 1. pp. 33-40.
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