Subcellular fractionation of polarized epithelial cells and identification of organelle-specific proteins by two-dimensional gel electrophoresis

Electrophoresis
I FialkaL A Huber

Abstract

Protein targeting and sorting is accomplished by complex vesicular transport processes that are tightly regulated within a cell. This is especially important for epithelial cells because correct delivery of newly synthesized proteins as well as recycling and sorting of internalized membrane proteins is essential for the establishment and preservation of cellular polarity. Many transport events, linking various subcellular compartments, have been analyzed, but many transport mechanisms still remain unresolved. In this study we attempted to identify proteins specifically associated with distinct organelles in murine mammary epithelial cells (EpH4). We isolated subcellular compartments by continuous sucrose gradient centrifugation in order to further analyze their protein composition by high-resolution two-dimensional gel electrophoresis (2-DE). The successful separation of late endosomes (LE), early endosomes (EE) and most of the rough endoplasmic reticulum (RER) was confirmed by subsequent analysis of gradient fractions for compartment-specific enzymes and marker proteins. Both Golgi and plasma membrane (PM) were found to partially co-purify with EE in such gradients. Characteristic polypeptide patterns were revealed on 2-DE gel...Continue Reading

References

Sep 1, 1992·The Journal of Cell Biology·A P HuovilaS D Fuller
Dec 1, 1989·The Journal of Cell Biology·G GriffithsM Marsh
Dec 1, 1989·The Journal of Cell Biology·M BomselK Simons
May 5, 1982·Journal of Molecular Biology·J Kyte, R F Doolittle
Aug 1, 1995·Current Opinion in Cell Biology·J Gruenberg, F R Maxfield
Jan 1, 1995·DNA Sequence : the Journal of DNA Sequencing and Mapping·A SaviozR W Davies
Feb 1, 1995·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·K E Mostov, M H Cardone
Aug 1, 1994·Current Opinion in Cell Biology·T Nilsson, G Warren
Jan 1, 1993·Journal of Cell Science. Supplement·S W Pimplikar, K Simons
Mar 1, 1993·The Journal of Cell Biology·N EmansJ Gruenberg
Feb 9, 1996·Cell·D G Drubin, W J Nelson
Feb 23, 1996·The Journal of Biological Chemistry·U LahtinenR F Pettersson
May 22, 1996·Biochimica Et Biophysica Acta·R H NewmanP Freemont
Jun 1, 1996·The Biochemical Journal·A PalmerE Knecht
Feb 1, 1996·Current Opinion in Immunology·P J Lehner, P Cresswell
Sep 1, 1996·Current Biology : CB·J M Lord
Dec 1, 1996·Molecular Biology of the Cell·S H LowK E Mostov
Dec 1, 1996·Molecular Biology of the Cell·H Y GaisanoW S Trimble
Jan 1, 1996·Annual Review of Cell and Developmental Biology·S R Pfeffer

❮ Previous
Next ❯

Citations

Nov 18, 2000·Electrophoresis·E JungD F Hochstrasser
Apr 4, 1998·Electrophoresis·C PasqualiL A Huber
Mar 23, 2010·The Journal of Membrane Biology·Keat-Eng NgAndrew Tinker
Jun 5, 2008·Journal of Neural Transmission·Ilse S PienaarJürgen Götz
Aug 5, 1999·Biochimie·V SantoniM Rossignol
Mar 6, 1999·Journal of Chromatography. B, Biomedical Sciences and Applications·C PasqualiL A Huber
Apr 11, 2013·Chemical Reviews·Chad P SatoriEdgar A Arriaga
Feb 26, 1999·The Plant Journal : for Cell and Molecular Biology·V SantoniM Rossignol
May 21, 2014·The Journal of Cell Biology·Natalia SchiefermeierLukas A Huber
Apr 15, 2014·Gastroenterology·Caroline L WiegerinckSabine Middendorp
Jul 5, 2005·Expert Opinion on Investigational Drugs·C R Stubberfield, M J Page
Nov 23, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Anna Bodzon-KulakowskaJerzy Silberring
Nov 13, 2004·Proteomics·Taras Stasyk, Lukas A Huber
Nov 13, 2004·Proteomics·Joerg ReindersAlbert Sickmann
Jul 27, 2007·Proteomics·Djuro Josic, James G Clifton
Jun 23, 2006·Proteomics·Sandra MorandellLukas A Huber
Nov 3, 2010·Mass Spectrometry Reviews·Ganesh Kumar AgrawalRandeep Rakwal
Feb 1, 2011·Proteomics·Sophie DuclosMichel Desjardins
Jul 9, 2004·Pathology, Research and Practice·Emanuela Felley-Bosco, Muriel André
Feb 13, 2007·Molecular & Cellular Proteomics : MCP·Taras StasykLukas A Huber
May 17, 2003·Circulation Research·Lukas A HuberIlja Vietor
Mar 9, 2002·Circulation Research·Mary F Lopez, Simon Melov
May 10, 2008·Journal of Cellular Biochemistry·Xiaochong WuJeffrey A Frost
Nov 17, 2010·Proteomics·Yie Hou LeeMaxey C M Chung
Nov 30, 2018·Current Protocols in Cell Biology·Daniel N ItzhakGeorg H H Borner

❮ Previous
Next ❯

Related Concepts

Related Feeds

Blood And Marrow Transplantation

The use of hematopoietic stem cell transplantation or blood and marrow transplantation (bmt) is on the increase worldwide. BMT is used to replace damaged or destroyed bone marrow with healthy bone marrow stem cells. Here is the latest research on bone and marrow transplantation.

B-Cell Lymphoma

B-cell lymphomas include lymphomas that affect B cells. This subtype of cancer accounts for over 80% of non-Hodgkin lymphomas in the US. Here is the latest research.