Association of prohormone convertase 3 with membrane lipid rafts

M. Blazquez, Kevin Docherty, Kathleen Isobel Joan Shennan

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Prohormone convertase 3 (PC3) is a neuroendocrine-specific member of the subtilisin-kexin family, involved in the intracellular processing and maturation of prohormones and proneuropeptides. PC3 is synthesised as a proprotein that undergoes two different cleavages resulting in the mature PC3 and the enzymatically active PC3 DeltaC. In vitro translated proPC3 and proPC3 DeltaC bind to trans-Golgi network (TGN)/granule-enriched membranes from the AtT20 neuroendocrine cell line in a pH-dependent manner suggesting both a dominant role for the pro-region in membrane association and that the C-terminal region is not essential. However, while PC3 bound to membranes the majority of PC3 DeltaC did not, suggesting that either the pro-region or the C-terminal region of PC3 is required for membrane association. Removal of peripheral membrane proteins did not affect the binding properties of any of the in vitro translated proteins. Chromaffin granule membranes (CGMs) were used to study the binding characteristics of endogenous PC3 and its active C-terminal truncated counterpart (PC3 DeltaC). Incubation of CGMs with Triton X-100 did not completely solubilise either of these forms of PC3. Moreover, both PC3 and PC3 DeltaC remained associated with detergent-resistant membrane microdomains, termed lipid rafts, purified from CGMs. The data raise the possibility that PC3 and PC3 DeltaC are sorted to the regulated secretory pathway via their association with membrane lipid rafts.

Original languageEnglish
Pages (from-to)107-116
Number of pages9
JournalJournal of Molecular Endocrinology
Volume27
Issue number1
DOIs
Publication statusPublished - 2001

Keywords

  • REGULATED SECRETORY PATHWAY
  • CARBOXYPEPTIDASE-E
  • ATT-20 CELLS
  • ATT20 CELLS
  • PC2
  • IDENTIFICATION
  • CLEAVAGE
  • DISTINCT
  • CDNA
  • BIOSYNTHESIS

Cite this

Association of prohormone convertase 3 with membrane lipid rafts. / Blazquez, M.; Docherty, Kevin; Shennan, Kathleen Isobel Joan.

In: Journal of Molecular Endocrinology, Vol. 27, No. 1, 2001, p. 107-116.

Research output: Contribution to journalArticle

@article{ef9a8b92f452411aa2b47016bcf4ab67,
title = "Association of prohormone convertase 3 with membrane lipid rafts",
abstract = "Prohormone convertase 3 (PC3) is a neuroendocrine-specific member of the subtilisin-kexin family, involved in the intracellular processing and maturation of prohormones and proneuropeptides. PC3 is synthesised as a proprotein that undergoes two different cleavages resulting in the mature PC3 and the enzymatically active PC3 DeltaC. In vitro translated proPC3 and proPC3 DeltaC bind to trans-Golgi network (TGN)/granule-enriched membranes from the AtT20 neuroendocrine cell line in a pH-dependent manner suggesting both a dominant role for the pro-region in membrane association and that the C-terminal region is not essential. However, while PC3 bound to membranes the majority of PC3 DeltaC did not, suggesting that either the pro-region or the C-terminal region of PC3 is required for membrane association. Removal of peripheral membrane proteins did not affect the binding properties of any of the in vitro translated proteins. Chromaffin granule membranes (CGMs) were used to study the binding characteristics of endogenous PC3 and its active C-terminal truncated counterpart (PC3 DeltaC). Incubation of CGMs with Triton X-100 did not completely solubilise either of these forms of PC3. Moreover, both PC3 and PC3 DeltaC remained associated with detergent-resistant membrane microdomains, termed lipid rafts, purified from CGMs. The data raise the possibility that PC3 and PC3 DeltaC are sorted to the regulated secretory pathway via their association with membrane lipid rafts.",
keywords = "REGULATED SECRETORY PATHWAY, CARBOXYPEPTIDASE-E, ATT-20 CELLS, ATT20 CELLS, PC2, IDENTIFICATION, CLEAVAGE, DISTINCT, CDNA, BIOSYNTHESIS",
author = "M. Blazquez and Kevin Docherty and Shennan, {Kathleen Isobel Joan}",
year = "2001",
doi = "10.1677/jme.0.0270107",
language = "English",
volume = "27",
pages = "107--116",
journal = "Journal of Molecular Endocrinology",
issn = "0952-5041",
publisher = "Society for Endocrinology",
number = "1",

}

TY - JOUR

T1 - Association of prohormone convertase 3 with membrane lipid rafts

AU - Blazquez, M.

AU - Docherty, Kevin

AU - Shennan, Kathleen Isobel Joan

PY - 2001

Y1 - 2001

N2 - Prohormone convertase 3 (PC3) is a neuroendocrine-specific member of the subtilisin-kexin family, involved in the intracellular processing and maturation of prohormones and proneuropeptides. PC3 is synthesised as a proprotein that undergoes two different cleavages resulting in the mature PC3 and the enzymatically active PC3 DeltaC. In vitro translated proPC3 and proPC3 DeltaC bind to trans-Golgi network (TGN)/granule-enriched membranes from the AtT20 neuroendocrine cell line in a pH-dependent manner suggesting both a dominant role for the pro-region in membrane association and that the C-terminal region is not essential. However, while PC3 bound to membranes the majority of PC3 DeltaC did not, suggesting that either the pro-region or the C-terminal region of PC3 is required for membrane association. Removal of peripheral membrane proteins did not affect the binding properties of any of the in vitro translated proteins. Chromaffin granule membranes (CGMs) were used to study the binding characteristics of endogenous PC3 and its active C-terminal truncated counterpart (PC3 DeltaC). Incubation of CGMs with Triton X-100 did not completely solubilise either of these forms of PC3. Moreover, both PC3 and PC3 DeltaC remained associated with detergent-resistant membrane microdomains, termed lipid rafts, purified from CGMs. The data raise the possibility that PC3 and PC3 DeltaC are sorted to the regulated secretory pathway via their association with membrane lipid rafts.

AB - Prohormone convertase 3 (PC3) is a neuroendocrine-specific member of the subtilisin-kexin family, involved in the intracellular processing and maturation of prohormones and proneuropeptides. PC3 is synthesised as a proprotein that undergoes two different cleavages resulting in the mature PC3 and the enzymatically active PC3 DeltaC. In vitro translated proPC3 and proPC3 DeltaC bind to trans-Golgi network (TGN)/granule-enriched membranes from the AtT20 neuroendocrine cell line in a pH-dependent manner suggesting both a dominant role for the pro-region in membrane association and that the C-terminal region is not essential. However, while PC3 bound to membranes the majority of PC3 DeltaC did not, suggesting that either the pro-region or the C-terminal region of PC3 is required for membrane association. Removal of peripheral membrane proteins did not affect the binding properties of any of the in vitro translated proteins. Chromaffin granule membranes (CGMs) were used to study the binding characteristics of endogenous PC3 and its active C-terminal truncated counterpart (PC3 DeltaC). Incubation of CGMs with Triton X-100 did not completely solubilise either of these forms of PC3. Moreover, both PC3 and PC3 DeltaC remained associated with detergent-resistant membrane microdomains, termed lipid rafts, purified from CGMs. The data raise the possibility that PC3 and PC3 DeltaC are sorted to the regulated secretory pathway via their association with membrane lipid rafts.

KW - REGULATED SECRETORY PATHWAY

KW - CARBOXYPEPTIDASE-E

KW - ATT-20 CELLS

KW - ATT20 CELLS

KW - PC2

KW - IDENTIFICATION

KW - CLEAVAGE

KW - DISTINCT

KW - CDNA

KW - BIOSYNTHESIS

U2 - 10.1677/jme.0.0270107

DO - 10.1677/jme.0.0270107

M3 - Article

VL - 27

SP - 107

EP - 116

JO - Journal of Molecular Endocrinology

JF - Journal of Molecular Endocrinology

SN - 0952-5041

IS - 1

ER -