Home > Research > Publications & Outputs > Ectodomain shedding of the Notch ligand Jagged1...

Links

Text available via DOI:

View graph of relations

Ectodomain shedding of the Notch ligand Jagged1 is mediated by ADAM17, but is not a lipid-raft-associated event

Research output: Contribution to Journal/MagazineJournal articlepeer-review

Published

Standard

Ectodomain shedding of the Notch ligand Jagged1 is mediated by ADAM17, but is not a lipid-raft-associated event. / Parr-Sturgess, Catherine A; Rushton, David J; Parkin, Edward T.
In: Biochemical Journal, Vol. 432, No. 2, 2010, p. 283-294.

Research output: Contribution to Journal/MagazineJournal articlepeer-review

Harvard

APA

Vancouver

Parr-Sturgess CA, Rushton DJ, Parkin ET. Ectodomain shedding of the Notch ligand Jagged1 is mediated by ADAM17, but is not a lipid-raft-associated event. Biochemical Journal. 2010;432(2):283-294. doi: 10.1042/BJ20100321

Author

Parr-Sturgess, Catherine A ; Rushton, David J ; Parkin, Edward T. / Ectodomain shedding of the Notch ligand Jagged1 is mediated by ADAM17, but is not a lipid-raft-associated event. In: Biochemical Journal. 2010 ; Vol. 432, No. 2. pp. 283-294.

Bibtex

@article{205143d5761f46b49573b48de6192137,
title = "Ectodomain shedding of the Notch ligand Jagged1 is mediated by ADAM17, but is not a lipid-raft-associated event",
abstract = "Notch signalling is an evolutionarily conserved pathway involved in cell-fate specification. The initiating event in this pathway is the binding of a Notch receptor to a DSL (Delta/Serrate/Lag-2) ligand on neighbouring cells triggering the proteolytic cleavage of Notch within its extracellular juxtamembrane region; a process known as proteolytic 'shedding' and catalysed by members of the ADAM (a disintegrin and metalloproteinase) family of enzymes. Jagged1 is a Notch-binding DSL ligand which is also shed by an ADAM-like activity raising the possibility of bi-directional cell-cell Notch signalling. In the present study we have unequivocally identified the sheddase responsible for shedding Jagged1 as ADAM17, the activity of which has previously been shown to be localized within specialized microdomains of the cell membrane known as 'lipid rafts'. However, we have shown that replacing the transmembrane and cytosolic regions of Jagged1 with a GPI (glycosylphosphatidylinositol) anchor, thereby targeting the protein to lipid rafts, did not enhance its shedding. Furthermore, the Jagged1 holoprotein, its ADAM-cleaved C-terminal fragment and ADAM17 were not enriched in raft preparations devoid of contaminating non-raft proteins. We have also demonstrated that wild-type Jagged1 and a truncated polypeptide-anchored variant lacking the cytosolic domain were subject to similar constitutive and phorbol ester-regulated shedding. Collectively these data demonstrate that Jagged1 is shed by ADAM17 in a lipid-raft-independent manner, and that the cytosolic domain of the former protein is not a pre-requisite for either constitutive or regulated shedding.",
keywords = "ADAM Proteins, Alkaline Phosphatase, Base Sequence, Calcium-Binding Proteins, Cell Membrane, Conserved Sequence, Cytosol, Evolution, Molecular, HEK293 Cells, Humans, Intercellular Signaling Peptides and Proteins, Membrane Microdomains, Membrane Proteins, Peptide Fragments, RNA, Small Interfering, Transfection",
author = "Parr-Sturgess, {Catherine A} and Rushton, {David J} and Parkin, {Edward T}",
year = "2010",
doi = "10.1042/BJ20100321",
language = "English",
volume = "432",
pages = "283--294",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "2",

}

RIS

TY - JOUR

T1 - Ectodomain shedding of the Notch ligand Jagged1 is mediated by ADAM17, but is not a lipid-raft-associated event

AU - Parr-Sturgess, Catherine A

AU - Rushton, David J

AU - Parkin, Edward T

PY - 2010

Y1 - 2010

N2 - Notch signalling is an evolutionarily conserved pathway involved in cell-fate specification. The initiating event in this pathway is the binding of a Notch receptor to a DSL (Delta/Serrate/Lag-2) ligand on neighbouring cells triggering the proteolytic cleavage of Notch within its extracellular juxtamembrane region; a process known as proteolytic 'shedding' and catalysed by members of the ADAM (a disintegrin and metalloproteinase) family of enzymes. Jagged1 is a Notch-binding DSL ligand which is also shed by an ADAM-like activity raising the possibility of bi-directional cell-cell Notch signalling. In the present study we have unequivocally identified the sheddase responsible for shedding Jagged1 as ADAM17, the activity of which has previously been shown to be localized within specialized microdomains of the cell membrane known as 'lipid rafts'. However, we have shown that replacing the transmembrane and cytosolic regions of Jagged1 with a GPI (glycosylphosphatidylinositol) anchor, thereby targeting the protein to lipid rafts, did not enhance its shedding. Furthermore, the Jagged1 holoprotein, its ADAM-cleaved C-terminal fragment and ADAM17 were not enriched in raft preparations devoid of contaminating non-raft proteins. We have also demonstrated that wild-type Jagged1 and a truncated polypeptide-anchored variant lacking the cytosolic domain were subject to similar constitutive and phorbol ester-regulated shedding. Collectively these data demonstrate that Jagged1 is shed by ADAM17 in a lipid-raft-independent manner, and that the cytosolic domain of the former protein is not a pre-requisite for either constitutive or regulated shedding.

AB - Notch signalling is an evolutionarily conserved pathway involved in cell-fate specification. The initiating event in this pathway is the binding of a Notch receptor to a DSL (Delta/Serrate/Lag-2) ligand on neighbouring cells triggering the proteolytic cleavage of Notch within its extracellular juxtamembrane region; a process known as proteolytic 'shedding' and catalysed by members of the ADAM (a disintegrin and metalloproteinase) family of enzymes. Jagged1 is a Notch-binding DSL ligand which is also shed by an ADAM-like activity raising the possibility of bi-directional cell-cell Notch signalling. In the present study we have unequivocally identified the sheddase responsible for shedding Jagged1 as ADAM17, the activity of which has previously been shown to be localized within specialized microdomains of the cell membrane known as 'lipid rafts'. However, we have shown that replacing the transmembrane and cytosolic regions of Jagged1 with a GPI (glycosylphosphatidylinositol) anchor, thereby targeting the protein to lipid rafts, did not enhance its shedding. Furthermore, the Jagged1 holoprotein, its ADAM-cleaved C-terminal fragment and ADAM17 were not enriched in raft preparations devoid of contaminating non-raft proteins. We have also demonstrated that wild-type Jagged1 and a truncated polypeptide-anchored variant lacking the cytosolic domain were subject to similar constitutive and phorbol ester-regulated shedding. Collectively these data demonstrate that Jagged1 is shed by ADAM17 in a lipid-raft-independent manner, and that the cytosolic domain of the former protein is not a pre-requisite for either constitutive or regulated shedding.

KW - ADAM Proteins

KW - Alkaline Phosphatase

KW - Base Sequence

KW - Calcium-Binding Proteins

KW - Cell Membrane

KW - Conserved Sequence

KW - Cytosol

KW - Evolution, Molecular

KW - HEK293 Cells

KW - Humans

KW - Intercellular Signaling Peptides and Proteins

KW - Membrane Microdomains

KW - Membrane Proteins

KW - Peptide Fragments

KW - RNA, Small Interfering

KW - Transfection

UR - http://www.scopus.com/inward/record.url?scp=78649583249&partnerID=8YFLogxK

U2 - 10.1042/BJ20100321

DO - 10.1042/BJ20100321

M3 - Journal article

C2 - 20819075

VL - 432

SP - 283

EP - 294

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 2

ER -