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PMID: 7575441 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Heterologous processing of rat prosomatostatin to somatostatin-14 by PC2: requirement for secretory cell but not the secretion granule.

The Biochemical journal ·Vol. 311 ( Pt 1) ·1995-10-01 ·Pages 111-8

Galanopoulou AS, Seidah NG, Patel YC

Abstract

The role of PC2 in prosomatostatin (PSS) processing was investigated in GH3/GH4C1 pituitary cells. These cells are sparsely granulated, express different amounts of PC2 and no PC1. We described heterologous processing of rat PSS (rPSS) co-expressed with PC2 in stably transfected cells, correlate PC2 protein levels under different conditions of transfection with efficiency of PSS processing to somatostatin-14 (SS-14), determine the effect of modulating cell granularity on enzyme expression and PSS processing, and compare the relative potency of PC2 with that of PC1, PSS and cleavage products were monitored by HPLC and radioimmunoassay of SS-like immunoreactivity (SSLI). Radioimmunoassay analysis of N-terminal PC2-like immunoreactivity (PC2 LI) in GH4C1:rPSS, GH4C1:rPSS + PC2 and GH3:rPSS transfectants showed a gradient of PC2 protein of 1:2.6:3.4 in cell extracts and 1:4.7:9 in secretion media from these cells respectively. The concentration of PC2 protein correlated with SS-14 conversion efficiency was 36 +/- 3% in GH4C1:rPSS cells, 56 +/- 7% in GH4C1:rPSS-PC2 cells and 100% in GH3:rPSS cells. Treatment of GH4C1:rPSS + PC2 cells with epidermal growth factor, insulin, and beta-estradiol to induce granules, significantly increased basal and forskolin-stimulated co-release of SS LI and PC2 LI, but had no influence on SS-14 processing efficiency. Hormone treatment led to a small increase in the ratio of mature PC2 (68 kDa) to proPC2 (75 kDa) forms. PC1 stably transfected in GH4C1 cells produced significantly greater SS-14 conversion (62% in cells, 66% in media) compared with PC2 transfectants (53% in cells, 47% in media) These results provide the first proof that PC2 can effect dibasic processing of mammalian PSS, and, along with PC1, qualifies as an authentic SS-14 convertase. The activity of PC2 requires the milieu of the secretory cell but not the secretory granule.

MeSH Terms
Animals Aspartic Acid Endopeptidases/genetics,metabolism Blotting, Northern Cell Line Colforsin/pharmacology Cytoplasmic Granules/metabolism Epidermal Growth Factor/pharmacology Estradiol/pharmacology Gene Expression Insulin/pharmacology Pituitary Gland/enzymology,metabolism,ultrastructure Proprotein Convertase 2 Proprotein Convertases Protein Precursors/genetics,metabolism RNA, Messenger/analysis,metabolism Rats Somatostatin/genetics,metabolism Subtilisins/genetics,metabolism Transfection
Chemicals
Insulin Protein Precursors RNA, Messenger Colforsin Estradiol Somatostatin Epidermal Growth Factor prosomatostatin Proprotein Convertases Subtilisins Proprotein Convertase 2 Aspartic Acid Endopeptidases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Galanopoulou A S
Fraser Laboratories, McGill University Department of Medicine, Royal Victoria Hospital, Montreal, Quebec, Canada.
Seidah N G
Patel Y C
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj3110111
Published
1995-10-01
Pages
111-8
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1136126
Subset
IM
External Links
PubMed source DOI: 10.1042/bj3110111 PMC Full Text View journal details
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