Organelle proteomics: implications for subcellular fractionation in proteomics.

Functional proteome analysis is not restricted to the sequence information but includes the broad spectrum of structural modifications and quantitative changes of proteins to which they are subjected in different tissues and cell organelles and during the development of an organism. Cell biology has provided the means required for the analysis of the composition and properties of purified cellular elements. Subcellular fractionation is an approach universal across all cell types and tissues, including cardiac and vascular system. Subcellular fractionation and proteomics form an ideal partnership when it comes to enrichment and analysis of intracellular organelles and low abundant multiprotein complexes. Subcellular fractionation is a flexible and adjustable approach resulting in reduced sample complexity and is most efficiently combined with high-resolution 2D gel/mass spectrometry analysis as well as with gel-independent techniques. In this study we introduce state of the art subcellular fractionation techniques and discuss their suitability, advantages, and limitations for proteomics research.

[1]  A. Görg,et al.  The current state of two‐dimensional electrophoresis with immobilized pH gradients , 2000, Electrophoresis.

[2]  L. Huber,et al.  Loss of epithelial polarity is accompanied by differential association of proteins with intracellular membranes , 1999, Electrophoresis.

[3]  S. Fleischer,et al.  Subcellular fractionation of rat liver. , 1974, Methods in enzymology.

[4]  E Gianazza,et al.  Immobilized pH gradients. , 1988, Trends in biochemical sciences.

[5]  M. Mann,et al.  Directed Proteomic Analysis of the Human Nucleolus , 2002, Current Biology.

[6]  T. Stevenson,et al.  Cardiac sarcoplasmic reticulum and sarcolemmal proteins separated by two‐dimensional electrophoresis: Surfactant effects on membrane solubilization , 2000, Electrophoresis.

[7]  Etienne Gagnon,et al.  Endoplasmic Reticulum-Mediated Phagocytosis Is a Mechanism of Entry into Macrophages , 2002, Cell.

[8]  D. K. Arrell,et al.  Proteomic Analysis of Pharmacologically Preconditioned Cardiomyocytes Reveals Novel Phosphorylation of Myosin Light Chain 1 , 2001, Circulation research.

[9]  H. Schwarz,et al.  Analysis of Cd44-Containing Lipid Rafts , 1999, The Journal of cell biology.

[10]  P. Steinlein,et al.  Identification of Syntenin as a Protein of the Apical Early Endocytic Compartment in Madin-Darby Canine Kidney Cells* , 1999, The Journal of Biological Chemistry.

[11]  K. Howell,et al.  Immuno-isolation of subcellular components. , 1988, Progress in clinical and biological research.

[12]  G. Griffiths,et al.  Gaining insight into a complex organelle, the phagosome, using two‐dimensional gel electrophoresis , 1995, Electrophoresis.

[13]  L. Huber,et al.  Subcellular fractionation, electromigration analysis and mapping of organelles. , 1999, Journal of chromatography. B, Biomedical sciences and applications.

[14]  A. Pfeifer,et al.  A Novel 14-Kilodalton Protein Interacts with the Mitogen-Activated Protein Kinase Scaffold Mp1 on a Late Endosomal/Lysosomal Compartment , 2001, The Journal of cell biology.

[15]  J. Perdue,et al.  An airfuge centrifugation procedure for the measurement of ligand binding to membrane-associated and detergent-solubilized plasma membrane receptors. , 1980, Journal of biochemical and biophysical methods.

[16]  Bradford W. Gibson,et al.  Characterization of the human heart mitochondrial proteome , 2003, Nature Biotechnology.

[17]  J. Gruenberg,et al.  Signaling from the far side. , 2002, Molecular cell.

[18]  G. Palade Cell fractionation: importance to cell-free systems development. , 1988, Progress in clinical and biological research.

[19]  S. Fleischer,et al.  Biochemical characterization of the golgi complex of mammalian cells. , 1974, Journal of supramolecular structure.

[20]  C. Pasquali,et al.  Preparative two‐dimensional gel electrophoresis of membrane proteins , 1997, Electrophoresis.

[21]  P. O’Farrell High resolution two-dimensional electrophoresis of proteins. , 1975, The Journal of biological chemistry.

[22]  S. Fleischer,et al.  [2] Subcellular fractionation of rat liver , 1974 .

[23]  P. Bork,et al.  Functional organization of the yeast proteome by systematic analysis of protein complexes , 2002, Nature.

[24]  M. Gerstein,et al.  A question of size: the eukaryotic proteome and the problems in defining it. , 2002, Nucleic acids research.

[25]  J. Strait,et al.  Isoenzyme-specific protein kinase C and c-Jun N-terminal kinase activation by electrically stimulated contraction of neonatal rat ventricular myocytes. , 2000, Journal of molecular and cellular cardiology.

[26]  K. Howell,et al.  Two‐dimensional mapping of the endogenous proteins of the rat hepatocyte Golgi complex cleared of proteins in transit , 1997, Electrophoresis.

[27]  J. Klose Protein mapping by combined isoelectric focusing and electrophoresis of mouse tissues , 1975, Humangenetik.

[28]  L. Huber,et al.  The recycling endosome of Madin-Darby canine kidney cells is a mildly acidic compartment rich in raft components. , 2000, Molecular biology of the cell.

[29]  K. Howell,et al.  Fusion in the endocytic pathway reconstituted in a cell-free system using immuno-isolated fractions. , 1988, Progress in clinical and biological research.

[30]  Y. Fujiki,et al.  Isolation of intracellular membranes by means of sodium carbonate treatment: application to endoplasmic reticulum , 1982, The Journal of cell biology.

[31]  S. Gygi,et al.  Quantitative analysis of complex protein mixtures using isotope-coded affinity tags , 1999, Nature Biotechnology.

[32]  K. Paiha,et al.  Both Calmodulin and the Unconventional Myosin Myr4 Regulate Membrane Trafficking Along the Recycling Pathway of MDCK Cells , 2000, Traffic.

[33]  A. Wandinger-Ness,et al.  Distinct transport vesicles mediate the delivery of plasma membrane proteins to the apical and basolateral domains of MDCK cells , 1990, The Journal of cell biology.

[34]  David S. Park,et al.  Caveolin-3 Knock-out Mice Develop a Progressive Cardiomyopathy and Show Hyperactivation of the p42/44 MAPK Cascade* , 2002, The Journal of Biological Chemistry.

[35]  L. Huber Is proteomics heading in the wrong direction? , 2003, Nature Reviews Molecular Cell Biology.

[36]  B. J. Hanson,et al.  A novel subfractionation approach for mitochondrial proteins: A three‐dimensional mitochondrial proteome map , 2001, Electrophoresis.

[37]  D. Teis,et al.  Localization of the MP1-MAPK scaffold complex to endosomes is mediated by p14 and required for signal transduction. , 2002, Developmental cell.

[38]  C. Brophy,et al.  Localization, Macromolecular Associations, and Function of the Small Heat Shock–Related Protein HSP20 in Rat Heart , 2003, Circulation.

[39]  K. Howell,et al.  Subcellular fractionation of tissue culture cells. , 1989, Trends in biochemical sciences.

[40]  S. Soboll,et al.  Cytosolic adenylates and adenosine release in perfused working heart. Comparison of whole tissue with cytosolic non-aqueous fractionation analyses. , 1986, European journal of biochemistry.

[41]  Etienne Gagnon,et al.  The Phagosome Proteome: Insight into Phagosome Functions , 2001 .

[42]  L. Huber,et al.  Subcellular fractionation of polarized epithelial cells and identification of organelle‐specific proteins by two‐dimensional gel electrophoresis , 1997, Electrophoresis.