Assays to measure PTEN lipid phosphatase activity in vitro from purified enzyme or immunoprecipitates

Laura Spinelli; Nicholas R. Leslie

doi:10.1007/978-1-4939-3746-2_6

Assays to measure PTEN lipid phosphatase activity in vitro from purified enzyme or immunoprecipitates

Laura Spinelli, Nicholas R. Leslie (Lead / Corresponding author)

Cell Signalling and Immunology

Research output: Chapter in Book/Report/Conference proceeding › Chapter (peer-reviewed) › peer-review

1 Citation (Scopus)

Abstract

PTEN is a one of the most frequently mutated tumor suppressors in human cancers. It is essential for regulating diverse biological processes and through its lipid phosphatase activity regulates the PI 3-Kinase signaling pathway. Sensitive phosphatase assays are employed to study the catalytic activity of PTEN against phospholipid substrates. Here we describe protocols to assay PTEN lipid phosphatase activity using either purified enzyme (purified PTEN lipid phosphatase assay) or PTEN immunopurified from tissues or cultured cells (cellular IP PTEN lipid phosphatase assay) against vesicles containing radiolabeled PIP3 substrate.

Original language	English
Title of host publication	Protein tyrosine phosphatases
Subtitle of host publication	methods and protocols
Editors	Rafael Pulido
Place of Publication	New York
Publisher	Springer
Pages	95-105
Number of pages	11
ISBN (Electronic)	9781493937462
ISBN (Print)	9781493937448
DOIs	https://doi.org/10.1007/978-1-4939-3746-2_6
Publication status	Published - 2016

Publication series

Name	Methods in molecular biology
Publisher	Springer
Volume	1447
ISSN (Print)	1064-3745

Keywords

Journal article
PTEN
Phosphoinositide
Phosphatase
Phosphate
Tumor suppressor
PI3-Kinase
Cancer

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1007/978-1-4939-3746-2_6

Aref#d: 21320. Antagonism of PI 3-Kinase Signalling by PTEN and SHIP2 (Programme Grant)
Downes, P. (Investigator) & Leslie, N. (Investigator)
Medical Research Council
1/10/09 → 30/09/14
Project: Research

Cite this

@inbook{1c0b4eec827a49219350b2812d6145bd,

title = "Assays to measure PTEN lipid phosphatase activity in vitro from purified enzyme or immunoprecipitates",

abstract = "PTEN is a one of the most frequently mutated tumor suppressors in human cancers. It is essential for regulating diverse biological processes and through its lipid phosphatase activity regulates the PI 3-Kinase signaling pathway. Sensitive phosphatase assays are employed to study the catalytic activity of PTEN against phospholipid substrates. Here we describe protocols to assay PTEN lipid phosphatase activity using either purified enzyme (purified PTEN lipid phosphatase assay) or PTEN immunopurified from tissues or cultured cells (cellular IP PTEN lipid phosphatase assay) against vesicles containing radiolabeled PIP3 substrate.",

keywords = "Journal article, PTEN, Phosphoinositide, Phosphatase, Phosphate, Tumor suppressor, PI3-Kinase, Cancer",

author = "Laura Spinelli and Leslie, {Nicholas R.}",

year = "2016",

doi = "10.1007/978-1-4939-3746-2_6",

language = "English",

isbn = "9781493937448",

series = "Methods in molecular biology",

publisher = "Springer ",

pages = "95--105",

editor = "Pulido, {Rafael }",

booktitle = "Protein tyrosine phosphatases",

}

Assays to measure PTEN lipid phosphatase activity in vitro from purified enzyme or immunoprecipitates. / Spinelli, Laura; Leslie, Nicholas R. (Lead / Corresponding author).
Protein tyrosine phosphatases: methods and protocols. ed. / Rafael Pulido. New York: Springer , 2016. p. 95-105 (Methods in molecular biology; Vol. 1447).

Research output: Chapter in Book/Report/Conference proceeding › Chapter (peer-reviewed) › peer-review

TY - CHAP

T1 - Assays to measure PTEN lipid phosphatase activity in vitro from purified enzyme or immunoprecipitates

AU - Spinelli, Laura

AU - Leslie, Nicholas R.

PY - 2016

Y1 - 2016

N2 - PTEN is a one of the most frequently mutated tumor suppressors in human cancers. It is essential for regulating diverse biological processes and through its lipid phosphatase activity regulates the PI 3-Kinase signaling pathway. Sensitive phosphatase assays are employed to study the catalytic activity of PTEN against phospholipid substrates. Here we describe protocols to assay PTEN lipid phosphatase activity using either purified enzyme (purified PTEN lipid phosphatase assay) or PTEN immunopurified from tissues or cultured cells (cellular IP PTEN lipid phosphatase assay) against vesicles containing radiolabeled PIP3 substrate.

AB - PTEN is a one of the most frequently mutated tumor suppressors in human cancers. It is essential for regulating diverse biological processes and through its lipid phosphatase activity regulates the PI 3-Kinase signaling pathway. Sensitive phosphatase assays are employed to study the catalytic activity of PTEN against phospholipid substrates. Here we describe protocols to assay PTEN lipid phosphatase activity using either purified enzyme (purified PTEN lipid phosphatase assay) or PTEN immunopurified from tissues or cultured cells (cellular IP PTEN lipid phosphatase assay) against vesicles containing radiolabeled PIP3 substrate.

KW - Journal article

KW - PTEN

KW - Phosphoinositide

KW - Phosphatase

KW - Phosphate

KW - Tumor suppressor

KW - PI3-Kinase

KW - Cancer

U2 - 10.1007/978-1-4939-3746-2_6

DO - 10.1007/978-1-4939-3746-2_6

M3 - Chapter (peer-reviewed)

C2 - 27514802

SN - 9781493937448

T3 - Methods in molecular biology

SP - 95

EP - 105

BT - Protein tyrosine phosphatases

A2 - Pulido, Rafael

PB - Springer

CY - New York

ER -

Assays to measure PTEN lipid phosphatase activity in vitro from purified enzyme or immunoprecipitates

Abstract

Publication series

Keywords

UN SDGs

Access to Document

Fingerprint

Projects

Aref#d: 21320. Antagonism of PI 3-Kinase Signalling by PTEN and SHIP2 (Programme Grant)

Cite this