ErbB2-associated changes in the lysosomal proteome

Jesper Nylandsted, Andrea C Becker, Jakob Bunkenborg, Jens S Andersen, Jörn Dengjel, Marja Jäättelä

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Late endosomes and lysosomes (hereafter referred to as lysosomes) play an essential role in the turnover of cellular macromolecules and organelles. Their biochemical characterization has so far depended on purification methods based on either density gradient centrifugations or magnetic purification of iron-loaded organelles. Owing to dramatic changes in lysosomal density and stability associated with lysosomal diseases and cancer, these methods are not optimal for the comparison of normal and pathological lysosomes. Here, we introduce an efficient method for the purification of intact lysosomes by magnetic immunoprecipitation with antibodies against the vacuolar-type H(+) -ATPase. Quantitative MS-based proteomics analysis of the obtained lysosomal membranes identified 60 proteins, most of which have previously been associated with the lysosomal compartment. Interestingly, the lysosomal membrane proteome was significantly altered by the ectopic expression of an active form of the ErbB2 oncogene, which renders the cells highly metastatic. The furthermost ErbB2-associated changes included increased levels of CD63, S100A11 and ferritin heavy chain. Overall, our data introduce the antibody-based purification of lysosomes as a suitable method for the characterization of lysosomes from a variety of pathological conditions with altered lysosomal density and stability.
OriginalsprogEngelsk
TidsskriftProteomics
Vol/bind11
Udgave nummer14
Sider (fra-til)2830-8
Antal sider9
ISSN1615-9853
DOI
StatusUdgivet - 1. jul. 2011

Fingeraftryk

Proteome
Purification
Apoferritins
Vacuolar Proton-Translocating ATPases
Membranes
Centrifugation
Antibodies
Macromolecules
Density Gradient Centrifugation
Iron
Proteins
Neoplasms

Citer dette

Nylandsted, J., Becker, A. C., Bunkenborg, J., Andersen, J. S., Dengjel, J., & Jäättelä, M. (2011). ErbB2-associated changes in the lysosomal proteome. Proteomics, 11(14), 2830-8. https://doi.org/10.1002/pmic.201000734
Nylandsted, Jesper ; Becker, Andrea C ; Bunkenborg, Jakob ; Andersen, Jens S ; Dengjel, Jörn ; Jäättelä, Marja. / ErbB2-associated changes in the lysosomal proteome. I: Proteomics. 2011 ; Bind 11, Nr. 14. s. 2830-8.
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Nylandsted, J, Becker, AC, Bunkenborg, J, Andersen, JS, Dengjel, J & Jäättelä, M 2011, 'ErbB2-associated changes in the lysosomal proteome', Proteomics, bind 11, nr. 14, s. 2830-8. https://doi.org/10.1002/pmic.201000734

ErbB2-associated changes in the lysosomal proteome. / Nylandsted, Jesper; Becker, Andrea C; Bunkenborg, Jakob; Andersen, Jens S; Dengjel, Jörn; Jäättelä, Marja.

I: Proteomics, Bind 11, Nr. 14, 01.07.2011, s. 2830-8.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - ErbB2-associated changes in the lysosomal proteome

AU - Nylandsted, Jesper

AU - Becker, Andrea C

AU - Bunkenborg, Jakob

AU - Andersen, Jens S

AU - Dengjel, Jörn

AU - Jäättelä, Marja

N1 - Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

PY - 2011/7/1

Y1 - 2011/7/1

N2 - Late endosomes and lysosomes (hereafter referred to as lysosomes) play an essential role in the turnover of cellular macromolecules and organelles. Their biochemical characterization has so far depended on purification methods based on either density gradient centrifugations or magnetic purification of iron-loaded organelles. Owing to dramatic changes in lysosomal density and stability associated with lysosomal diseases and cancer, these methods are not optimal for the comparison of normal and pathological lysosomes. Here, we introduce an efficient method for the purification of intact lysosomes by magnetic immunoprecipitation with antibodies against the vacuolar-type H(+) -ATPase. Quantitative MS-based proteomics analysis of the obtained lysosomal membranes identified 60 proteins, most of which have previously been associated with the lysosomal compartment. Interestingly, the lysosomal membrane proteome was significantly altered by the ectopic expression of an active form of the ErbB2 oncogene, which renders the cells highly metastatic. The furthermost ErbB2-associated changes included increased levels of CD63, S100A11 and ferritin heavy chain. Overall, our data introduce the antibody-based purification of lysosomes as a suitable method for the characterization of lysosomes from a variety of pathological conditions with altered lysosomal density and stability.

AB - Late endosomes and lysosomes (hereafter referred to as lysosomes) play an essential role in the turnover of cellular macromolecules and organelles. Their biochemical characterization has so far depended on purification methods based on either density gradient centrifugations or magnetic purification of iron-loaded organelles. Owing to dramatic changes in lysosomal density and stability associated with lysosomal diseases and cancer, these methods are not optimal for the comparison of normal and pathological lysosomes. Here, we introduce an efficient method for the purification of intact lysosomes by magnetic immunoprecipitation with antibodies against the vacuolar-type H(+) -ATPase. Quantitative MS-based proteomics analysis of the obtained lysosomal membranes identified 60 proteins, most of which have previously been associated with the lysosomal compartment. Interestingly, the lysosomal membrane proteome was significantly altered by the ectopic expression of an active form of the ErbB2 oncogene, which renders the cells highly metastatic. The furthermost ErbB2-associated changes included increased levels of CD63, S100A11 and ferritin heavy chain. Overall, our data introduce the antibody-based purification of lysosomes as a suitable method for the characterization of lysosomes from a variety of pathological conditions with altered lysosomal density and stability.

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DO - 10.1002/pmic.201000734

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JO - Proteomics

JF - Proteomics

SN - 1615-9853

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Nylandsted J, Becker AC, Bunkenborg J, Andersen JS, Dengjel J, Jäättelä M. ErbB2-associated changes in the lysosomal proteome. Proteomics. 2011 jul 1;11(14):2830-8. https://doi.org/10.1002/pmic.201000734