Mapping the protein composition of trans-Golgi network (TGN)-derived carrier vesicles from polarized MDCK cells

Electrophoresis
K FiedlerK Simons

Abstract

In polarized MDCK cells, proteins and lipids are sorted in the trans-Golgi network /TGN) and packaged into different vesicular carriers that are delivered to the apical or basolateral cell surface. To gain insight into the sorting and trafficking machinery, we have previously isolated TGN-derived carrier vesicles from perforated MDCK cells. The composition of immuno-isolated apical and basolateral carriers was mapped by two-dimensional (2-D) gel electrophoresis. Here we describe the identification of several components of the vesicle fraction by using three different methods. 2-D gel comigration was performed with carrier vesicles isolated from metabolically labeled MDCK cells and human epidermal keratinocyte lysates. This allowed us to assign eleven known components by a comparison with the comprehensive keratinocyte 2-D gel database. These comprised two members of the 14-3-3 family of proteins that have been implicated in vesicular trafficking. Five proteins were purified from preparative 2-D gels and identified by peptide microsequencing, including the beta1 and beta2 subunit of trimeric G proteins and an annexin II variant. A member of the SNARE family of proteins was identified by immunoblotting. The combination of 2-D gel...Continue Reading

References

Oct 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·E Rodriguez Boulan, D D Sabatini
Aug 15, 1975·European Journal of Biochemistry·R A Laskey, A D Mills
Sep 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·K GalvinM Ozturk
Aug 1, 1992·Current Opinion in Cell Biology·J Gruenberg, M J Clague
Jan 1, 1992·Annual Review of Cell Biology·E Rodriguez-Boulan, S K Powell
Feb 21, 1992·Cell·K G RothbergR G Anderson
Nov 1, 1991·Electrophoresis·H H RasmussenJ Vandekerckhove
Jul 27, 1990·Cell·K Simons, A Wandinger-Ness
Oct 5, 1990·Journal of Molecular Biology·S F AltschulD J Lipman
Aug 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·R G MakiP K Srivastava
Jan 1, 1986·Journal of Biochemistry·S Ohta, Y Kagawa
Sep 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·S Nakajima-IijimaT Kakunaga
Jul 10, 1987·Nucleic Acids Research·B Dworniczak, M E Mirault
Aug 23, 1988·Biochemistry·K Simons, G van Meer
Jun 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·H K FongM I Simon
Jan 1, 1985·Annual Review of Cell Biology·K Simons, S D Fuller
May 1, 1984·Journal of Biochemical and Biophysical Methods·P Tunón, K E Johansson
Nov 2, 1995·Nature·P ScheiffeleK Simons
May 1, 1995·Trends in Biochemical Sciences·K Fiedler, K Simons
May 5, 1995·Cell·K Fiedler, K Simons
Nov 3, 1994·Nature·J E Rothman
Apr 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·M K Bennett, R H Scheller
Jun 6, 1994·FEBS Letters·T V KurzchaliaS Monier
Aug 1, 1993·Current Opinion in Cell Biology·M Zerial, H Stenmark
Nov 1, 1993·Electrophoresis·J E CelisM Nielsen
Jul 15, 1993·The Biochemical Journal·R D Burgoyne, A Morgan

❮ Previous
Next ❯

Citations

Nov 18, 2000·Electrophoresis·E JungD F Hochstrasser
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
Mar 2, 2010·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·Asne R BauskinSamuel N Breit
Nov 4, 2004·BMC Cell Biology·Edward L OrganDonald H Rubin
Sep 16, 1998·Proceedings of the National Academy of Sciences of the United States of America·R BeisswangerW B Huttner
Nov 17, 2006·American Journal of Physiology. Renal Physiology·Michael G JanechJohn M Arthur
Aug 5, 2003·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Giuseppe ProcinoGiovanna Valenti
Jul 5, 2003·Physiological Reviews·Pamela L Tuma, Ann L Hubbard

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.