Electromigration for separations of protein complexes.

This paper describes electromigration of complexes, consisting of two or more proteins and non-covalently associated peptides. Relatively small complexes (Mr < 1000000) can be resolved in sieving matrices. Large complexes are separated in free liquid systems. Examples of separation are given using native gels, denaturing gels and special formats thereof: blue native PAGE and gels incorporating a transversal temperature gradient. Both preparative and analytical applications are discussed as well as separations leading to mechanistic models of protein interaction. Carrier-free electrophoresis is represented by capillary zone electrophoresis, free-flow electrophoresis and density gradient electrophoresis. Emphasis is put on the free liquid separation of clathrin-coated vesicles and proteasomes.

[1]  K. Hendil,et al.  Human proteasomes analysed with monoclonal antibodies. , 1995, The Biochemical journal.

[2]  M. Rechsteiner,et al.  Multiple forms of the 20 S multicatalytic and the 26 S ubiquitin/ATP-dependent proteases from rabbit reticulocyte lysate. , 1992, The Journal of biological chemistry.

[3]  W. Schlote,et al.  Human Diseases with Defects in Oxidative Phosphorylation , 1995 .

[4]  B. Karger,et al.  Affinity probe capillary electrophoresis: analysis of recombinant human growth hormone with a fluorescent labeled antibody fragment. , 1994, Analytical chemistry.

[5]  R. Christopherson,et al.  Analysis of the binding of deoxyribonuclease I to G‐actin by capillary electrophoresis , 1997, Electrophoresis.

[6]  K. Jalink,et al.  High performance density gradient electrophoresis of subcellular organelles, protein complexes and proteins , 1998, Electrophoresis.

[7]  H. Schägger,et al.  Human diseases with defects in oxidative phosphorylation. 2. F1F0 ATP-synthase defects in Alzheimer disease revealed by blue native polyacrylamide gel electrophoresis. , 1995, European journal of biochemistry.

[8]  J. Drijfhout,et al.  Mannose receptor‐mediated uptake of antigens strongly enhances HLA class II‐restricted antigen presentation by cultured dendritic cells , 1997, European journal of immunology.

[9]  John Calvin Reed,et al.  Drug-induced apoptosis in B-cell chronic lymphocytic leukemia: relationship between p53 gene mutation and bcl-2/bax proteins in drug resistance. , 1996, Oncogene.

[10]  H. Schägger.  Quantification of oxidative phosphorylation enzymes after blue native electrophoresis and two‐dimensional resolution: Normal complex I protein amounts in Parkinson's disease conflict with reduced catalytic activities , 1995, Electrophoresis.

[11]  P. Kloetzel,et al.  Maturation of mammalian 20 S proteasome: purification and characterization of 13 S and 16 S proteasome precursor complexes. , 1997, Journal of molecular biology.

[12]  I. Krull,et al.  Capillary isoelectric focusing as a tool in the examination of antibodies, peptides and proteins of pharmaceutical interest. , 1996, Journal of chromatography. A.

[13]  A. Chrambach,et al.  Preparative electrophoresis in “sieving media” of subcellular‐sized particles , 1998, Electrophoresis.

[14]  J. Pieters,et al.  Application of an improved density gradient electrophoresis apparatus to the separation of proteins, cells and subcellular organelles , 1993, Electrophoresis.

[15]  A C Steven,et al.  Applicability of agarose gel electrophoresis to the physical characterization of clathrin-coated vesicles. , 1985, Analytical biochemistry.

[16]  J. Rothman,et al.  Purification of coated vesicles by agarose gel electrophoresis , 1981, The Journal of cell biology.

[17]  H. Braun,et al.  New insights into the composition, molecular mass and stoichiometry of the protein complexes of plant mitochondria. , 1996, The Plant journal : for cell and molecular biology.

[18]  N. Kedersha,et al.  Subpopulations of liver coated vesicles resolved by preparative agarose gel electrophoresis , 1986, The Journal of cell biology.

[19]  H. Ploegh,et al.  Intracellular interactions of transferrin and its receptor during biosynthesis , 1990, The Journal of cell biology.

[20]  E. Ungewickell,et al.  Auxilin, a newly identified clathrin-associated protein in coated vesicles from bovine brain , 1990, The Journal of cell biology.

[21]  P. Cresswell,et al.  In vivo and in vitro formation and dissociation of HLA-DR complexes with invariant chain-derived peptides. , 1994, Immunity.

[22]  J. Neefjes,et al.  Direct vesicular transport of MHC class II molecules from lysosomal structures to the cell surface , 1996, The Journal of cell biology.

[23]  D. Carey,et al.  Self-association of N-Syndecan (Syndecan-3) Core Protein Is Mediated by a Novel Structural Motif in the Transmembrane Domain and Ectodomain Flanking Region (*) , 1995, The Journal of Biological Chemistry.

[24]  D. Rickwood,et al.  Gel electrophoresis of proteins: a practical approach , 1981 .

[25]  H. Schägger.  Native electrophoresis for isolation of mitochondrial oxidative phosphorylation protein complexes. , 1995, Methods in enzymology.

[26]  B. Oudega,et al.  The Escherichia coli K88 periplasmic chaperone FaeE forms a heterotrimeric complex with the minor fimbrial component FaeH and with the minor fimbrial component FaeI. , 1995, Microbial pathogenesis.

[27]  R. G. Nielsen,et al.  Separation of antibody-antigen complexes by capillary zone electrophoresis, isoelectric focusing and high-performance size-exclusion chromatography. , 1991, Journal of chromatography.

[28]  J. Neefjes,et al.  High resolution density gradient electrophoresis of cellular organelles , 1996, Electrophoresis.

[29]  M. Robinson,et al.  The role of clathrin, adaptors and dynamin in endocytosis. , 1994, Current opinion in cell biology.

[30]  H. Ploegh,et al.  Isolation and characterization of the intracellular MHC class II compartment , 1994, Nature.

[31]  M. Yu,et al.  Refolding of alpha 1-antitrypsin expressed as inclusion bodies in Escherichia coli: characterization of aggregation. , 1995, Biochimica et biophysica acta.

[32]  D. Verwoerd,et al.  Analysis of members of the Ig‐gene superfamily by thermal gradient polyacrylamide gel electrophoresis , 1992, Electrophoresis.

[33]  A. Chrambach,et al.  Towards predicting mobility and resolution in polymeric media: Some first steps , 1998, Electrophoresis.

[34]  H. Schägger,et al.  Blue native electrophoresis for isolation of membrane protein complexes in enzymatically active form. , 1991, Analytical biochemistry.

[35]  K. Tsuji.  Evaluation of sodium dodecyl sulfate non-acrylamide, polymer gel-filled capillary electrophoresis for molecular size separation of recombinant bovine somatotropin. , 1993, Journal of chromatography. A.

[36]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[37]  B. Barber,et al.  A recommendation for visualizing disulfide bonding by one-dimensional sodium dodecyl sulfate--polyacrylamide gel electrophoresis. , 1984, Analytical biochemistry.

[38]  H. Schägger,et al.  Analysis of molecular masses and oligomeric states of protein complexes by blue native electrophoresis and isolation of membrane protein complexes by two-dimensional native electrophoresis. , 1994, Analytical biochemistry.

[39]  D. Verwoerd,et al.  Thermostability analysis of major histocompatibility complex class I molecules by temperature gradient gel electrophoresis , 1993, European journal of immunology.

[40]  A. Rivett,et al.  Subpopulations of proteasomes in rat liver nuclei, microsomes and cytosol. , 1996, The Biochemical journal.

[41]  K. Pfeiffer,et al.  Electrophoretic separation of multiprotein complexes from blood platelets and cell lines: Technique for the analysis of diseases with defects in oxidative phosphorylation , 1996, Electrophoresis.

[42]  M. Schlaak,et al.  Conventional isoelectric focusing does not dissociate immune complexes. , 1987, Journal of immunological methods.

[43]  K. von Figura,et al.  Lysosomal enzyme precursors in coated vesicles derived from the exocytic and endocytic pathways , 1987, The Journal of cell biology.

[44]  J. Neefjes,et al.  High‐resolution density gradient electrophoresis of proteins and subcellular organelles , 1997, Electrophoresis.