Hermann Schägger

article

Large pore gels to separate mega protein complexes larger than 10 MDa by blue native electrophoresis: Isolation of putative respiratory strings or patches

Valentina Strecker, Zibiernisha Wumaier, Ilka Wittig, Hermann Schägger

Here, we expand the application of blue native electrophoresis to the separation of mega protein complexes larger than 10 MDa by introducing novel large pore acrylamide gels. We tailored the bis‐acrylamide cross‐linker amounts relative to the acrylamide monomer to enlarge the pore size of acrylamide gels and to obtain elastic and sufficiently stable gels. The novel gel types were then used to search...

article

Isolation of putative respiratory strings or patches by blue-native electrophoresis on large pore gels

Valentina Strecker, Zibiernisha Wumaier, Ilka Wittig, Hermann Schägger

BBA - Bioenergetics > 2010 > 1797 > Supplement > 73

article

Novel applications of blue-native PAGE

Ilka Wittig, Zibiernisha Wumaier, Valentina Strecker, Hermann Schägger

BBA - Bioenergetics > 2010 > 1797 > Supplement > 71

article

Assembly and oligomerization of human ATP synthase lacking mitochondrial subunits a and A6L

Ilka Wittig, Bjoern Meyer, Heinrich Heide, Mirco Steger, more

BBA - Bioenergetics > 2010 > 1797 > 6-7 > 1004-1011

Here we study ATP synthase from human ρ ⁰ (rho zero) cells by clear native electrophoresis (CNE or CN-PAGE) and show that ATP synthase is almost fully assembled in spite of the absence of subunits a and A6L. This identifies subunits a and A6L as two of the last subunits to complete the ATP synthase assembly. Minor amounts of dimeric and even tetrameric forms of the large assembly intermediate...

article

Laser‐induced liquid bead ion desorption‐MS of protein complexes from blue‐native gels, a sensitive top‐down proteomic approach

Lucie Sokolova, Ilka Wittig, Hans‐Dieter Barth, Hermann Schägger, more

PROTEOMICS > 10 > 7 > 1401 - 1407

We have developed an experimental approach that combines two powerful methods for proteomic analysis of large membrane protein complexes: blue native electrophoresis (BNE or BN‐PAGE) and laser‐induced liquid bead ion desorption (LILBID) MS. Protein complexes were separated by BNE and eluted from the gel. The masses of the constituents of the multiprotein complexes were obtained by LILBID MS, a detergent‐tolerant...

article

Native immunoblotting of blue native gels to identify conformation‐specific antibodies

Volker Zickermann, Zibiernisha Wumaier, Blanka Wrzesniewska, Carola Hunte, more

PROTEOMICS > 10 > 1 > 159 - 163

A large repertoire of immunological methods permits monitoring the interaction of antibodies with their specific antigen. However, recognition of a protein by a conformation‐specific antibody represents a challenge because native conditions must be kept throughout the assay. Native immunoblotting of blue native gels conserves the native state by using Tween 20 instead of methanol for the obligatory...

article

Mitochondrial ABC proteins in health and disease

Ariane Zutz, Simone Gompf, Hermann Schägger, Robert Tampé

BBA - Bioenergetics > 2009 > 1787 > 6 > 681-690

ABC transporters represent one of the largest families of membrane proteins that are found in all three phyla of life. Mitochondria comprise up to four ABC systems, ABCB7/ATM1, ABCB10/MDL1, ABCB8 and ABCB6. These half-transporters, which assemble into homodimeric complexes, are involved in a number of key cellular processes, e.g. biogenesis of cytosolic iron–sulfur clusters, heme biosynthesis, iron...

article

Role of phospholipids in respiratory cytochrome bc1 complex catalysis and supercomplex formation

Tina Wenz, Ruth Hielscher, Petra Hellwig, Hermann Schägger, more

BBA - Bioenergetics > 2009 > 1787 > 6 > 609-616

Specific protein–lipid interactions have been identified in X-ray structures of membrane proteins. The role of specifically bound lipid molecules in protein function remains elusive. In the current study, we investigated how phospholipids influence catalytic, spectral and electrochemical properties of the yeast respiratory cytochrome bc ₁ complex and how disruption of a specific cardiolipin...

article

Supramolecular organization of ATP synthase and respiratory chain in mitochondrial membranes

Ilka Wittig, Hermann Schägger

BBA - Bioenergetics > 2009 > 1787 > 6 > 672-680

Mitochondrial ATP synthase is mostly isolated in monomeric form, but in the inner mitochondrial membrane it seems to dimerize and to form higher oligomeric structures from dimeric building blocks. Following a period of electron microscopic single particle analyses that revealed an angular orientation of the membrane parts of monomeric ATP synthases in the dimeric structures, and after extensive studies...

article

S1/6 Structural organization of mitochondrial ATP synthase

Ilka Wittig, Hermann Schägger

BBA - Bioenergetics > 2008 > 1777 > Supplement > S10

article

Structural organization of mitochondrial ATP synthase

Ilka Wittig, Hermann Schägger

BBA - Bioenergetics > 2008 > 1777 > 7-8 > 592-598

Specific modules and subcomplexes like F ₁ and F ₀ -parts, F ₁ -c subcomplexes, peripheral and central stalks, and the rotor part comprising a ring of c-subunits with attached subunits γ, δ, and ε can be identified in yeast and mammalian ATP synthase. Four subunits, α ₃ β ₃ , OSCP, and h, seem to form a structural entity at the extramembranous rotor/stator interface...

article

Supercomplexes and subcomplexes of mitochondrial oxidative phosphorylation

Ilka Wittig, Rosalba Carrozzo, Filippo M. Santorelli, Hermann Schägger

BBA - Bioenergetics > 2006 > 1757 > 9-10 > 1066-1072

Dimerization or oligomerization of ATP synthase has been proposed to play an important role for mitochondrial cristae formation and to be involved in regulating ATP synthase activity. We found comparable oligomycin-sensitive ATPase activity for monomeric and oligomeric ATP synthase suggesting that oligomerization/monomerization dynamics are not directly involved in regulating ATP synthase activity...

article

Temporary attachment of carbohydrates to cyclopeptide templates: a new strategy for single-bead analysis of multivalent neoglycopeptides

Valentin Wittmann, Sonja Seeberger, Hermann Schägger

Tetrahedron Letters > 2003 > 44 > 52 > 9243 - 9246

Employing a cleavable carbohydrate-peptide linker, a new strategy for single-bead analysis of multivalent cyclic neoglycopeptides based on Edman degradation is described. Edman degradation of glycopeptides is hampered by the acid lability of the glycosidic bond and potential incompatibilities of phenylthiohydantoin (PTH) derivatives of glycosylated amino acids with PTH derivatives of the proteinogenic...

article

Redox-Induced Transitions in Bovine Cytochrome bc ₁ Complex Studied by Perfusion-Induced ATR-FTIR Spectroscopy ^†

Masayo Iwaki, Livia Giotta, Akinyemi O. Akinsiku, Hermann Schägger, more

Biochemistry > 2003 > 42 > 38 > 11109 - 11119

article

Isolation, Characterization and Electron Microscopic Single Particle Analysis of the NADH:Ubiquinone Oxidoreductase (Complex I) from the Hyperthermophilic Eubacterium Aquifex aeolicus ^†

Guohong Peng, Günter Fritzsch, Volker Zickermann, Hermann Schägger, more

Biochemistry > 2003 > 42 > 10 > 3032 - 3039

article

Structural Insights into the A ₁ ATPase from the Archaeon, Methanosarcina mazei Gö1 ^†

Gerhard Grüber, Dmitri I. Svergun, Ünal Coskun, Thorsten Lemker, more

Biochemistry > 2001 > 40 > 7 > 1890 - 1896

article

Biophysical and structural characterization of proton-translocating NADH-dehydrogenase (complex I) from the strictly aerobic yeast Yarrowia lipolytica

Rogieh Djafarzadeh, Stefan Kerscher, Klaus Zwicker, Michael Radermacher, more

BBA - Bioenergetics > 2000 > 1459 > 1 > 230-238

Mitochondrial proton-translocating NADH-dehydrogenase (complex I) is one of the largest and most complicated membrane bound protein complexes. Despite its central role in eukaryotic oxidative phosphorylation and its involvement in a broad range of human disorders, little is known about its structure and function. Therefore, we have started to use the powerful genetic tools available for the strictly...

article

Binding of detergents and inhibitors to bovine complex I - a novel purification procedure for bovine complex I retaining full inhibitor sensitivity

Jürgen G. Okun, Volker Zickermann, Klaus Zwicker, Hermann Schägger, more

BBA - Bioenergetics > 2000 > 1459 > 1 > 77-87

Mitochondrial complex I exhibits some peculiar and poorly understood features regarding the effects of detergents on activity and sensitivity to hydrophobic inhibitors that are not seen with other membrane complexes using ubiquinone as a substrate. Therefore, we investigated the interaction of complex I from bovine heart mitochondria with different types of detergents by monitoring activity, degree...

article

Chromaffin granule membrane glycoprotein IV is identical with Ac45, a membrane-integral subunit of the granule's H ⁺ -ATPase

Fariha Getlawi, Andrea Laslop, Hermann Schägger, Jürgen Ludwig, more

Neuroscience Letters > 1996 > 219 > 1 > 13-16

Glycoprotein IV of bovine adrenal chromaffin granule membranes was purified by membrane fractionation with Triton X-114 and lectin affinity chromatography. An antiserum raised against this protein recognized the same component as one directed against subunit Ac45 of the proton-translocating adenosine triphosphatase in the granule membrane. Amino acid sequencing confirmed that glycoprotein IV and...

article

Multiple deficiencies of mitochondrial DNA- and nuclear-encoded subunits of respiratory NADH dehydrogenase detected with peptide- and subunit-specific antibodies in mitochondrial myopathies

Herman A.C.M. Bentlage, Antoon J.M. Janssen, Anne Chomyn, Giuseppe Attardi, more

BBA - Biomembranes > 1995 > 1234 > 1 > 63-73

Antibodies have been raised against synthetic peptides corresponding to several computer-predicted epitopes of three mtDNA-encoded subunits, ND4, ND5 and ND6, of the human respiratory chain NADH dehydrogenase (Complex I). Antibodies were characterized by a sensitive immunoblotting assay using proteins from human skeletal muscle mitochondria and by immunoprecipitation of radio-labeled HeLa cell mitochondrial...

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Search results for: Hermann Schägger

Large pore gels to separate mega protein complexes larger than 10 MDa by blue native electrophoresis: Isolation of putative respiratory strings or patches

Isolation of putative respiratory strings or patches by blue-native electrophoresis on large pore gels

Novel applications of blue-native PAGE

Assembly and oligomerization of human ATP synthase lacking mitochondrial subunits a and A6L

Laser‐induced liquid bead ion desorption‐MS of protein complexes from blue‐native gels, a sensitive top‐down proteomic approach

Native immunoblotting of blue native gels to identify conformation‐specific antibodies

Mitochondrial ABC proteins in health and disease

Role of phospholipids in respiratory cytochrome bc1 complex catalysis and supercomplex formation

Supramolecular organization of ATP synthase and respiratory chain in mitochondrial membranes

S1/6 Structural organization of mitochondrial ATP synthase

Structural organization of mitochondrial ATP synthase

Supercomplexes and subcomplexes of mitochondrial oxidative phosphorylation

Temporary attachment of carbohydrates to cyclopeptide templates: a new strategy for single-bead analysis of multivalent neoglycopeptides

Redox-Induced Transitions in Bovine Cytochrome bc ₁ Complex Studied by Perfusion-Induced ATR-FTIR Spectroscopy ^†

Isolation, Characterization and Electron Microscopic Single Particle Analysis of the NADH:Ubiquinone Oxidoreductase (Complex I) from the Hyperthermophilic Eubacterium Aquifex aeolicus ^†

Structural Insights into the A ₁ ATPase from the Archaeon, Methanosarcina mazei Gö1 ^†

Biophysical and structural characterization of proton-translocating NADH-dehydrogenase (complex I) from the strictly aerobic yeast Yarrowia lipolytica

Binding of detergents and inhibitors to bovine complex I - a novel purification procedure for bovine complex I retaining full inhibitor sensitivity

Chromaffin granule membrane glycoprotein IV is identical with Ac45, a membrane-integral subunit of the granule's H ⁺ -ATPase

Multiple deficiencies of mitochondrial DNA- and nuclear-encoded subunits of respiratory NADH dehydrogenase detected with peptide- and subunit-specific antibodies in mitochondrial myopathies

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