A proteomics analysis of rat liver membrane skeletons: the investigation of actin- and cytokeratin-based protein components.

Membrane skeletons, which are defined for their resistance to Triton extraction of cell membrane, play a pivotal role in cell shape and signal transduction. In the present work, we applied a complementary proteomics strategy: 2-DE combined with MALDI-TOF MS and 1D-PAGE coupled with LC-ESI-FTICR MS to analyze a membrane skeleton fraction isolated from Sprague-Dawley (SD) rat livers. We report confident identification of 104 proteins (39 membrane skeleton proteins) using 2-DE and MALDI-TOF MS approach and 402 proteins (87 membrane skeleton proteins) using 1D-PAGE LC-MS/MS analysis. In total, 100 membrane skeleton proteins were identified using the two complementary proteomics means. To the best of our knowledge, this is the largest data set of membrane skeleton proteins to date. Noteworthily, almost all of these membrane skeleton proteins were associated with actin or cytokeratin, and more than half of them were involved in various cell junctions. Our results offer insights into the protein components of the actin- and cytokeratin-based membrane skeletons in rat livers, which would improve our understanding of their biological roles.

[1]  E. Feener,et al.  Label-free Quantitative Analysis of One-dimensional PAGE LC/MS/MS Proteome , 2008, Molecular & Cellular Proteomics.

[2]  Lingyao Meng,et al.  Comparative Studies of Early Liver Dysfunction in Senescence-accelerated Mouse Using Mitochondrial Proteomics Approaches*S , 2008, Molecular & Cellular Proteomics.

[3]  L. Rothfield,et al.  Duplication and segregation of the actin (MreB) cytoskeleton during the prokaryotic cell cycle , 2007, Proceedings of the National Academy of Sciences.

[4]  Qingsong Wang,et al.  Proteomic analysis of a membrane skeleton fraction from human liver. , 2007, Journal of proteome research.

[5]  Carole Yee,et al.  A proteomic characterization of the plasma membrane of human epidermis by high-throughput mass spectrometry. , 2004, The Journal of investigative dermatology.

[6]  L. Kedes,et al.  Cytoplasmic Nuclear Transfer of the Actin-capping Protein Tropomodulin* , 2004, Journal of Biological Chemistry.

[7]  M. Mann,et al.  Trypsin Cleaves Exclusively C-terminal to Arginine and Lysine Residues*S , 2004, Molecular & Cellular Proteomics.

[8]  Ruedi Aebersold,et al.  Complementary Analysis of the Mycobacterium tuberculosis Proteome by Two-dimensional Electrophoresis and Isotope-coded Affinity Tag Technology * , 2004, Molecular & Cellular Proteomics.

[9]  J. Yates,et al.  The application of mass spectrometry to membrane proteomics , 2003, Nature Biotechnology.

[10]  J. Leszyk,et al.  Proteomic Analysis of a Detergent-resistant Membrane Skeleton from Neutrophil Plasma Membranes* 210 , 2002, The Journal of Biological Chemistry.

[11]  E. Nordhoff,et al.  Alpha-cyano-4-hydroxycinnamic acid affinity sample preparation. A protocol for MALDI-MS peptide analysis in proteomics. , 2001, Analytical chemistry.

[12]  J. Yates,et al.  Direct analysis of protein complexes using mass spectrometry , 1999, Nature Biotechnology.

[13]  A. Shevchenko,et al.  Femtomole sequencing of proteins from polyacrylamide gels by nano-electrospray mass spectrometry , 1996, Nature.

[14]  B. Burke,et al.  Cytoskeleton-membrane interactions. , 1996, Current opinion in cell biology.

[15]  E J Luna,et al.  Cytoskeleton--plasma membrane interactions. , 1992, Science.

[16]  P. Bayley,et al.  Localization of two fibronectin-binding glycoproteins in rat liver and primary hepatocytes. Co-distribution in vitro of integrin (alpha 5 beta 1) and non-integrin (AGp110) receptors in cell-substratum adhesion sites. , 1990, Journal of cell science.

[17]  S. Tsukita,et al.  Isolation of cell-to-cell adherens junctions from rat liver , 1989, The Journal of cell biology.

[18]  J. Glenney,et al.  Fodrin is the general spectrin-like protein found in most cells whereas spectrin and the TW protein have a restricted distribution , 1983, Cell.

[19]  K. Willecke,et al.  Immunocytochemical localization of the gap junction 26 K protein in mouse liver plasma membranes. , 1983, The EMBO journal.

[20]  D. Wall,et al.  Isolation of rat hepatocyte plasma membranes. I. Presence of the three major domains , 1983, The Journal of cell biology.

[21]  A. Hubbard,et al.  Isolation of rat hepatocyte plasma membranes. II. Identification of membrane-associated cytoskeletal proteins , 1983, The Journal of cell biology.

[22]  Vann Bennett,et al.  Brain spectrin, a membrane-associated protein related in structure and function to erythrocyte spectrin , 1982, Nature.

[23]  L. Hood,et al.  Rat liver gap junction protein: properties and partial sequence. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[24]  L. Orci,et al.  In vivo induction of tight junction proliferation in rat liver , 1976, The Journal of cell biology.

[25]  D. Friend,et al.  HIGH-YIELD PREPARATION OF ISOLATED RAT LIVER PARENCHYMAL CELLS , 1969, Journal of Cell Biology.

[26]  G. Palade,et al.  JUNCTIONAL COMPLEXES IN VARIOUS EPITHELIA , 1963, The Journal of cell biology.

[27]  R. Flaumenhaft,et al.  The actin cytoskeleton differentially regulates platelet alpha-granule and dense-granule secretion. , 2005, Blood.

[28]  大西 弘恵.  JACOP, a novel plaque protein localizing at the apical junctional complex with sequence similarity to cingulin , 2005 .

[29]  E A Khairallah,et al.  Differential detergent fractionation of isolated hepatocytes: Biochemical, immunochemical and two‐dimensional gel electrophoresis characterization of cytoskeletal and noncytoskeletal compartments , 1994, Electrophoresis.

[30]  T. Steck,et al.  Selective solubilization of proteins and phospholipids from red blood cell membranes by nonionic detergents. , 1973, Journal of supramolecular structure.