Blue native PAGE

Nature Protocols

Volumn 1, Issue 1, 2006, Pages 418-428

  Wittig, Ilka ^a   Braun, Hans Peter ^b   Schägger, Hermann ^a  

^a UNIVERSITY HOSPITAL FRANKFURT   (Germany)

^b LEIBNIZ UNIVERSITY HANNOVER   (Germany)

Author keywords

[No Author keywords available]

Indexed keywords

MEMBRANE PROTEIN;

ARTICLE; ISOELECTRIC FOCUSING; ISOLATION AND PURIFICATION; METABOLISM; METHODOLOGY; MITOCHONDRIAL MEMBRANE; POLYACRYLAMIDE GEL ELECTROPHORESIS;

ELECTROPHORESIS, POLYACRYLAMIDE GEL; ISOELECTRIC FOCUSING; MEMBRANE PROTEINS; MITOCHONDRIAL MEMBRANES;

EID: 33750378958     PISSN: 17542189     EISSN: 17502799     Source Type: Journal    
DOI: 10.1038/nprot.2006.62     Document Type: Article

References (55)

1. Schaägger, H. & von Jagow, G. Blue native electrophoresis for isolation of membrane protein complexes in enzymatically active form. Anal. Biochem. 199, 223-231 (1991).
2. Schaägger, H., Cramer, W.A. & von Jagow, G. 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. Anal. Biochem. 217, 220-230 (1994).
3. Schaägger, H. Blue native electrophoresis, in Membrane Protein Purification and Crystallization. A Practical Guide 2nd edn. (eds. Hunte, C., von Jagow, G. & Schägger, H.) 5.105-5.130 (Academic, San Diego, USA, 2003).
4. Kügler, M., Jänsch, L., Kruft, V., Schmitz, U.K. & Braun, H.P. Analysis of the chloroplast protein complexes by blue-native polyacrylamide gelelectrophoresis. Photosynth. Res. 53, 35-44 (1997).
5. Schägger, H. Blue native gels to isolate protein complexes from mitochondria. Methods Cell Biol. 65, 231-244 (2001).
6. Pfeiffer, K. et al. Cardiolipin stabilizes respiratory chain supercomplexes. J. Biol. Chem. 278, 52873-52880 (2003).
7. Ludwig, J. et al. Identification and characterization of a novel 9.2 kDa membrane sector associated protein of vacuolar proton-ATPase from chromaffin granules. J. Biol. Chem. 273, 10939-10947 (1998).
8. Schägger, H. Native electrophoresis for isolation of mitochondrial oxidative phosphorylation protein complexes. Methods Enzymol. 260, 190-202 (1995).
9. Vahsen, N. et al. AIF deficiency compromises oxidative phosphorylation. EMBO J. 23, 4679-4689 (2004).
10. Acin-Perez, R. et al. Respiratory complex III is required to maintain complex I in mammalian mitochondria. Mol. Cell 13, 805-815 (2004).
11. Schägger, H. et al. Significance of respirasomes for the assembly/stability of human respiratory chain complex I. J. Biol. Chem. 279, 36349-36353 (2004).
12. Camacho-Carvajal, M., Wollscheid, B., Aebersold, R., Steimle, V. & Schamel, W.A. Two-dimensional blue native/SDS gel electrophoresis of multi-protein complexes from whole cellular lysates. Mol. Cell. Proteomics 3, 176-182 (2004).
13. Schägger, H. et al. Electrophoretic separation of multiprotein complexes from blood platelets and cell lines: Technique for the analysis of diseases with defects in oxidative phosphorylation. Electrophoresis 17, 709-714 (1996).
14. Schägger, H. Quantification of oxidative phosphorylation enzymes after blue native electrophoresis and two-dimensional resolution: Normal complex I protein amounts in Parkinsońs disease conflict with reduced catalytic activities. Electrophoresis 16, 763-770 (1995).
15. Carrozzo, R. et al. Subcomplexes of human ATP synthase mark mitochondrial biosynthesis disorders. Ann. Neurol. 59, 265-275 (2006).
16. Dudkina, N.V., Heinemeyer, J., Keegstra, W., Boekema, E.J. & Braun, H.P. Structure of dimeric ATP synthase from mitochondria: An angular association of monomers induces the strong curvature of the inner membrane. FEBS Lett. 579, 5769-5772 (2005).
17. Dudkina, N.V., Eubel, H., Keegstra, W., Boekema, E.J. & Braun, H.P. Structure of a mitochondrial supercomplex formed by respiratory-chain complexes I and III. Proc. Natl. Acad. Sci. USA 102, 3225-3229 (2005).
18. 0 bridging features and the structural basis of mitochondrial cristae biogenesis. Proc. Natl. Acad. Sci. USA 102, 12356-12358 (2005).
19. Schamel, W.W. et al. Coexistence of multivalent and monovalent TCRs explains high sensitivity and wide range of response. J. Exp. Med. 202, 493-503 (2005).
20. 0-ATP synthase exists as a dimer: identification of three dimer-specific subunits. EMBO J. 17, 7170-7178 (1998).
21. Schägger, H. & Pfeiffer, K. Supercomplexes in the respiratory chains of yeast and mammalian mitochondria. EMBO J. 19, 1777-1783 (2000).
22. Stroh, A. et al. Assembly of respiratory chain complexes I, III, and IV into NADH oxidase supercomplex stabilizes Complex I in Paracoccus denitrificans. J. Biol. Chem. 279, 5000-5007 (2004).
23. Eubel, H., Jänsch, L. & Braun, H.P. New insights into the respiratory chain of plant mitochondria: Supercomplexes and a unique composition of complex II. Plant Physiol. 133, 274-286 (2003).
24. Eubel, H., Heinemeyer, J. & Braun, H.P. Identification and characterization of respirasomes in potato mitochondria. Plant Physiol. 134, 1450-1459 (2004).
25. Paumard, P. et al. The ATP synthase is involved in generating mitochondrial cristae morphology. EMBO J. 21, 221-230 (2002).
26. Poetsch, A., Neff, D., Seelert, H., Schägger, H. & Dencher, N.A. Dye removal, catalytic activity and 2D-crystallization of chloroplast H+-ATP synthase purified by blue native electrophoresis. Biochim. Biophys. Acta 1466, 339-349 (2000).
27. Zerbetto, E., Vergani, L. & Dabbeni-Sala, F. Quantification of muscle mitochondrial oxidative phosphorylation enzymes via histochemical staining of blue native polyacrylamide gels. Electrophoresis 18 2059-2064 (1997).
28. Jung, C., Higgins, C.M.J. & Xu, Z. Measuring the quantity of mitochondrial electron transport chain complexes in tissues of central nervous system using Blue native polyacrylamide gel electrophoresis. Anal. Biochem. 286, 214-223 (2000).
29. Eubel, H., Heinemeyer, J., Sunderhaus, S. & Braun, H.P. Respiratory chain supercomplexes in plant mitochondria. Plant Physiol. Biochem. 42, 937-942 (2004).
30. Sunderhaus, S. et al. Carbonic anhydrase subunits form a matrix-exposed domain attached to the membrane arm of mitochondrial complex I in plants. J. Biol. Chem. 281, 6482-6488 (2006).
31. Griffon, N. et al. Molecular determinants of glycine receptor subunit assembly. EMBO J. 18, 4711-4721 (1999).
32. Dietmeyer, K. et al. Tom5 functionally links mitochondrial preprotein receptors to the general import pore. Nature 388, 195-200 (1997).
33. Jänsch, L., Kruft, V., Schmitz, U.K. & Braun, H.P. Unique composition of the preprotein translocase of the outer mitochondrial membrane from plants. J. Biol. Chem. 273, 17251-17257 (1998).
34. Rais, I., Karas, M. & Schägger, H. Two-dimensional electrophoresis for the isolation of integral membrane proteins and mass spectrometric identification. Proteomics 4, 2567-2571 (2004).
35. Fandino, A.S. et al. LC-nanospray-MS/MS analysis of hydrophobic proteins from membrane protein complexes isolated by blue-native electrophoresis. J. Mass Spectrom. 40, 1223-1231 (2005).
36. Perales, M. et al. Disruption of a nuclear gene encoding a mitochondrial gamma carbonic anhydrase reduces complex I and supercomplex I+III2 levels and alters mitochondrial physiology in Arabidopsis. J. Mol. Biol. 350, 263-277 (2005).
37. Wittig, I. & Schägger, H. Advantages and limitations of clear native polyacrylamide gel electrophoresis. Proteomics 5, 4338-4346 (2005).
38. 0-ATP synthase. Biochim. Biophys. Acta 1607, 167-179 (2003).
39. 0-ATP synthase complex interactions in vivo can occur in the absence of the dimer specific subunit e. J. Bioenerg. Biomembr. 37, 55-66 (2005).
40. Schägger, H. & von Jagow, G. Tricine-sodium dodecyl sulfate polyacrylamide gel electrophoresis for the separation of proteins in the range from 1-100 kDalton. Anal. Biochem. 166, 368-379 (1987).
41. Schägger, H. SDS electrophoresis techniques, in Membrane Protein Purification and Crystallization. A Practical Guide 2nd edn. (eds. Hunte, C., von Jagow, G. & Schägger, H.) 4.85-4.103 (Academic, San Diego, USA, 2003).
42. Schägger, H. Tricine-SDS-PAGE. Nat. Protocols 1, 16-22 (2006).
43. Werhahn, W. & Braun, H.P. Biochemical dissection of the mitochondrial proteome from Arabidopsis thaliana by three-dimensional gel electrophoresis. Electrophoresis 23, 640-646 (2002).
44. Studier, F.W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J. Mol. Biol. 79, 237-248 (1973).
45. Hunkapiller, M.W., Lujan, E., Ostrander, F. & Hood, L.E. Isolation of microgram quantities of proteins from polyacrylamide gels for amino acid sequence analysis. Methods Enzymol. 91, 227-236 (1983).
46. Hjerten, S. Chromatographic separation according to size of macromolecules and cell particles on columns of agarose suspensions. Arch. Biochem. Biophys. 99, 466-475 (1962).
47. Reif, S., Voos, W. & Rassow, J. Intramitochondrial dimerization of citrate synthase characterized by blue native electrophoresis. Anal. Biochem. 288, 97-99 (2001).
48. Meisinger, C., Pfanner, N. & Truscott, K.N. Isolation of yeast mitochondria. Methods Mol. Biol. 313, 33-39 (2006).
49. Heinemeyer, J., Eubel, H., Wehmhöhner, D., Jänsch, L. & Braun, H.P. Proteomic approach to characterize the supramolecular organization of photosystems in higher plants. Phytochemistry 65, 1683-1692 (2004).
50. Klose, J. & Kobalz, U. Two-dimensional electrophoresis of proteins: An updated protocol and implications for a functional analysis of the genome. Electrophoresis 16, 1034-1059 (1995).
51. Görg, A. & Weiss, W., High-resolution two-dimensional electrophoresis of proteins using immobilized pH gradients. in Cell Biology. A Laboratory Handbook (ed. Celis, J.) 386-397 (Academic, New York, 1998).
52. Laemmli, U.K. Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227, 680-685 (1970).
53. Schägger, H., Brandt, U., Gencic, S. & von Jagow, G. Ubiquinol-cytochrome c-reductase from human and bovine mitochondria. Methods Enzymol. 260, 82-96 (1995).
54. Moro, F., Sirrenberg, C., Schneider, H.C., Neupert, W. & Brunner, M. The TIM17.23 preprotein translocase of mitochondria: Composition and function in protein transport into the matrix. EMBO J. 18, 3667-3675 (1999).
55. Schamel, W.W. & Reth, M. Monomeric and oligomeric complexes of the B cell antigen receptor. Immunity 13, 5-14 (2000).