DOCSLIB.ORG

Imported Mitochondrial Proteins Cytochrome B2 And

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Imported Mitochondrial Proteins Cytochrome B2 And Proc. NatL Acad. Sci. USA Vol. 79, pp. 267-271, January 1982 Biochemistry Imported mitochondrial proteins cytochrome b2 and cytochrome cl are processed in two steps (mitochondrial biogenesis/protein import/proteolytic processing/in vitro protein synthesis/vectorial processing) SUSAN M. GASSER, AKIRA OHASHI, GUNTHER DAUM, PETER C. BOHNI, JANE GIBSON, GRAEME A. REID, TAKASHILYONETANI, AND GOTTFRIED SCHATZ Department of Biochemistry, Biocenter, University of Basel, CH-4056, Basel, Switzerland Communicated by Tadeus Reichstein, September 14, 1981 ABSTRACT Cytochrome b2 of yeast is located in the space 1 2 3 4 5 6 between the inner and outer mitochondrial membranes whereas cytochrome cl is bound to the outer face ofthe mitochondrial inner membrane. Both proteins are made outside the mitochondria as larger precursors that are processed to their mature forms in two pre b, steps. In the first step, at least a segment of the precursor poly- cyt b2 - peptide chain penetrates into the mitochondrial matrix and is cleaved to an intermediate form by a matrix-localized soluble pro- tease. This step requires an electrochemical gradient across the mitochondrial inner membrane. In the second step, the inter- mediate form is converted to the mature form. For cytochrome cl, this second step requires heme. Import of these two cyto- chromes thus involves a "detour" across the inner membrane. Most mitochondrial proteins are synthesized outside the mi- are than the cor- tochondria as precursors that usually larger FIG. 1. Import of cytochrome b2 into isolated yeast mitochondria responding mature polypeptides found inside the mitochondria is dependent on ATP. Total yeast mRNA was translated in a reticu- (1, 2). Very little is known about how these precursors are trans- locyte lysate in the presence of [35S]methionine (13), centrifuged for ported into the mitochondria. A promising experimental system 40 min at 140,000 x g to remove ribosomes, and filtered through a for studying this question is the uptake ofradiolabeled in vitro- Sephadex G-25 column equilibrated with 0.15 M KCI/20 mM Hepes- synthesized precursors by isolated mitochondria. In vitro up- KOH, pH 7.4. Aliquots (150 AD) of the filtered lysate were incubated take does not require protein synthesis and, with larger pre- with isolated yeast mitochondria (200 ,ug of protein; total vol, 0.4 ml) for 30 min at 280C. Samples in lanes 1 and 2 also contained 1 mM ATP/ cursors, is accompanied by conversion of the precursor to its 1 mM MgCl2/5 mM phosphoenolpyruvate and 4 units of pyruvate ki- mature size ("vectorial processing"; ref. 1). We have isolated nase. Samples in lanes 1, 2, 5, and 6 were subsequently treated for 30 a soluble chelator-sensitive protease from the yeast mitochon- min at 00C with 150 pg of trypsin/ml and then with 3 mg of soybean drial matrix that processes precursors of those proteins trans- trypsin inhibitor and 1 mM tosyl-L-lysyl-chlormethyl ketone. Mito- ported partly or completely across the inner membrane (3). chondria were then sedimented (10 min at 10,000 x g) and pellets these to the corresponding mature (lanes 1, 3, and 5) and supernatants (lanes 2,4, and 6) were dissociated Processing of precursors ir>3% NaDodSO4and subjectedto immunoprecipitation (13) with anti- polypeptides appears to occur in a single step. serum against cytochrome b2. The immunoprecipitated samples were Here we report that cytochrome b2, a soluble enzyme ofthe analyzed by NaDodSO/10% polyacrylamide gel electrophoresis and mitochondrial intermembrane space, is processed to its mature fluorography. A photograph of the fluorogram is shown. pre b2, Cy- size in two steps. Surprisingly, the first ofthese steps occurs in tochrome b2 precursor; cyt b2, mature cytochrome b2. the matrix space. Cytochrome cl, an inner membrane enzyme the space, is processed by a protruding into intermembrane cytochrome cl) or 10% (for cytochrome b2) gel slabs (8). Pub- similar two-step mechanism. lished methods were used for assaying protein (9), fumarase (10), and L-lactate-ferricyanide reductase (11); for fluorography METHODS ofdried gel slabs (12); and for isolating crude yeast mRNA (13), Growth and Labeling of Cells. Wild-type Saccharomyces spheroplasts (7), yeast mitochondria (14), soluble processing cerevisiae strain D273-10B (ATCC 25657) was grown on semi- protease from a yeast mitochondrial matrix (3), and yeast cy- synthetic medium (4) containing 0.3% galactose (unless stated tochrome cl (15). The isolation of subfractions from yeast mi- otherwise). The heme-deficient mutant GL-1 (5) was grown on tochondria will be described in detail elsewhere but details are semisynthetic medium containing 0.26% Tween 80, 12 ppm of available on request. ergosterol, and 2% glucose. Uniform labeling of cells with Immunological Methods. A monoclonal antibody against 3So2- or [3S]methionine (4) and pulse labeling of intact cells subunit III ofyeast cytochrome oxidase was raised by injecting (6) or spheroplasts (7) have been described. holocytochrome c oxidase into BALB/c mice and fusing spleen Analytical and Preparative Methods. NaDodSO4poly- lymphocytes with X63-Ag8 myeloma cells (16). Published meth- acrylamide gel electrophoresis was carried out on 12.5% (for ods were used for raising antisera in rabbits (17), for immuno- precipitation ofcell fractions dissociated in NaDodSO4 (13), and The publication costs ofthis article were defrayed in part by page charge for immune replication (18). payment. This article must therefore be hereby marked "advertise- Cell-Free Translation. Rabbit reticulocyte lysates were ment" in accordance with 18 U. S. C. §1734 solely to indicate this fact. mied with the necessary cofactors (19), frozen in liquid N2, and 267 Downloaded by guest on September 27, 2021 268 Biochemistry: Gasser et al. Proc. Natl. Acad. Sci. USA 79 (1982) Table 1. Import of cytochrome b2 into isolated mitochondria 1 2 3 4 Mitochondria Supernatant Trypsin absent 50 42* Trypsin present 47 <1 _ _pre b. Results are relative to amount of precursor immunoprecipitated cvt b.) from lysate before incubation (defined as 100%). Isolated yeast mito- chondria were allowed to import in vitro-synthesized precursor of cy- tochrome b2 in the presence of ATP as described in Fig. 1. To ensure equal efficiency of immunoprecipitation from the different samples, their contents of unlabeled cytochrome b2 were equalized by adding 200 ,g of unlabeled yeast mitochondria to each supernatant and to an aliquot of the labeled reticulocyte lysate that served as a 100% control. The samples were then dissociated with NaDodSO4 and treated as in Fig. 1. The fluorograph was quantitated with a Bausch and Lomb den- sitometer. Results refer to mature cytochrome b2 except where indi- cated otherwise. FIG. 2. ATP-dependent import of cytochrome b2 into isolated mi- * 31% precursor/11% mature. tochondria is blocked by carboxyatractylate. The experiment was sim- ilar to that described in Fig. 1 except that all samples contained the ATP mixture; trypsin treatment was omitted; and during incubation stored at -80'C without nuclease treatment. Theywere labeled ofthe mitochondria withlysate, 20 pg ofcarboxyatractylate was added with [3S]methionine [=100 Ci/mmol; 10 mCi/ml (1 Ci = 3.7 to the samples in lanes 3 and 4. Lanes: 1 and 3, pellet; 2 and 4, super- x 1010 becquerels); prepared according to ref. 20 from natant, pre b2, Cytochrome b2 precursor; cyt b2, mature cytochrome b2. 35SO2-] in the presence of 10-15 OD260 units of crude yeast mRNA per ml for 60 min at 29°C and treated further as outlined duced in vitro by incubating isolated yeast mitochondria with in Fig. 1. in vitro-synthesized cytochrome b2 precursor in the presence Materials. Yeast cytochrome b2 was a gift of F. Lederer and ofATP. Under these conditions, 50% ofthe labeled cytochrome F. S. Matthews. A rabbit antiserum against yeast isopropyl- b2 precursor is taken up by the mitochondria and converted to malate synthase was donated by P. Roeder and G. Kohlhaw. mature cytochrome b2 (Fig. 1), which is almost completely re- All other chemicals were purchased. sistant to externally added trypsin (Table 1). In the absence of ATP, the precursor is neither taken up nor processed (Fig. 1). RESULTS Vectorial processing of precursor polypeptides by isolated mi- Cytochrome b2 (L-lactate-cytochrome c reductase) of yeast is tochondria is thus energy dependent. This confirms the earlier coded in the nucleus, synthesized outside the mitochondria, observation that vectorial processing ofmitochondrial precursor and imported into the organelles (21). The mature enzyme is polypeptides in yeast spheroplasts requires energy (7). a soluble component of the mitochondrial intermembrane ATP-dependent import and processing ofcytochrome b2 are space. This location is documented by the following observa- blocked ifentry of the added ATP into the mitochondrial matrix tions (unpublished data): (i) on subfractionating yeast mito- is prevented by carboxyatractylate (Fig. 2). chondria, cytochrome b2 cofractionates with adenylate kinase Addition ofoligomycin (which blocks mitochondrial ATPase, (an intermembrane space marker), but not with fumarase, acon- ref. 22) or ofvalinomycin/K+ (which results in a collapse ofthe itase, or citrate synthase (three matrix markers); (ii) in intact membrane potential, ref. 22) also completely inhibits the im- mitochondria, the enzyme readily reduces added ferricyanide port of cytochrome b2 into mitochondria. However, if mito- (which can penetrate the outer but not the inner membrane) chondria are energized with a respiratory substrate such as suc- but fails to reduce added cytochrome c (which cannot penetrate cinate, instead ofwith ATP, oligomycin does not inhibit import either of the two membranes); and (iii) an antiserum against (not shown). Import of precursors into mitochondria thus re- cytochrome b2 inhibits the mitochondria-bound enzyme only quires an electrochemical gradient across the mitochondrial in- if the outer membrane is disrupted. ner membrane. The mitochondrial outer membrane cannot sus- Import of cytochrome b2 into mitochondria can be repro- tain such a gradient because it permits diffusion of molecules Table 2.
Load more

Recommended publications
  • Establishing the Pathogenicity of Novel Mitochondrial DNA Sequence Variations: a Cell and Molecular Biology Approach
    Mafalda Rita Avó Bacalhau Establishing the Pathogenicity of Novel Mitochondrial DNA Sequence Variations: a Cell and Molecular Biology Approach Tese de doutoramento do Programa de Doutoramento em Ciências da Saúde, ramo de Ciências Biomédicas, orientada pela Professora Doutora Maria Manuela Monteiro Grazina e co-orientada pelo Professor Doutor Henrique Manuel Paixão dos Santos Girão e pela Professora Doutora Lee-Jun C. Wong e apresentada à Faculdade de Medicina da Universidade de Coimbra Julho 2017 Faculty of Medicine Establishing the pathogenicity of novel mitochondrial DNA sequence variations: a cell and molecular biology approach Mafalda Rita Avó Bacalhau Tese de doutoramento do programa em Ciências da Saúde, ramo de Ciências Biomédicas, realizada sob a orientação científica da Professora Doutora Maria Manuela Monteiro Grazina; e co-orientação do Professor Doutor Henrique Manuel Paixão dos Santos Girão e da Professora Doutora Lee-Jun C. Wong, apresentada à Faculdade de Medicina da Universidade de Coimbra. Julho, 2017 Copyright© Mafalda Bacalhau e Manuela Grazina, 2017 Esta cópia da tese é fornecida na condição de que quem a consulta reconhece que os direitos de autor são pertença do autor da tese e do orientador científico e que nenhuma citação ou informação obtida a partir dela pode ser publicada sem a referência apropriada e autorização. This copy of the thesis has been supplied on the condition that anyone who consults it recognizes that its copyright belongs to its author and scientific supervisor and that no quotation from the
    [Show full text]
  • Electron Transport Discovery Four Complexes Complex I: Nadhà Coqh2
    BI/CH 422/622 OUTLINE: Pyruvate pyruvate dehydrogenase Krebs’ Cycle How did he figure it out? Overview 8 Steps Citrate Synthase Aconitase Isocitrate dehydrogenase Ketoglutarate dehydrogenase Succinyl-CoA synthetase Succinate dehydrogenase Fumarase Malate dehydrogenase Energetics Regulation Summary Oxidative Phosphorylation Energetics (–0.16 V needed for making ATP) Mitochondria Transport (2.4 kcal/mol needed to transport H+ out) Electron transport Discovery Four Complexes Complex I: NADHà CoQH2 Complex II: Succinateà CoQH2 2+ Complex III: CoQH2à Cytochrome C (Fe ) 2+ Complex IV: Cytochrome C (Fe ) à H2O Electron Transport à O2 Inhibitors of Electron Transport Big Drop! • Inhibitors all stop ET and ATP synthesis: very toxic! Spectral work Big Drop! NADH Cyto-a3 Cyto-c1 Big Drop! Cyto-b Cyto-c Cyto-a Fully reduced Flavin Cyto-c + rotenone + antimycin A 300 350 400 450 500 550 600 650 700 1 Electron Transport Electron-Transport Chain Complexes Contain a Series of Electron Carriers • Better techniques for isolating and handling mitochondria, and isolated various fractions of the inner mitochondrial membrane • Measure E°’ • They corresponded to these large drops, and they contained the redox compounds isolated previously. • When assayed for what reactions they could perform, they could perform certain redox reactions and not others. • When isolated, including isolating the individual redox compounds, and measuring the E°’ for each, it was clear that an electron chain was occurring; like a wire! • Lastly, when certain inhibitors were added, some of the redox reactions could be inhibited and others not. Site of the inhibition could be mapped. Electron Transport Electron-Transport Chain Complexes Contain a Series of Electron Carriers • Better techniques for isolating and handling mitochondria, and isolated various fractions of the inner mitochondrial membrane • Measure E°’ • They corresponded to these large drops, and they contained the redox compounds isolated previously.
    [Show full text]
  • Function and Structure of Complex Ii of The
    Annu. Rev. Biochem. 2003. 72:77–109 doi: 10.1146/annurev.biochem.72.121801.161700 FUNCTION AND STRUCTURE OF COMPLEX II * OF THE RESPIRATORY CHAIN Gary Cecchini Molecular Biology Division, Veterans Administration Medical Center, San Francisco, California 94121, and Department of Biochemistry and Biophysics, University of California, San Francisco, California 94143; email: [email protected] Key Words succinate dehydrogenase, fumarate reductase, quinone oxidoreductase, reactive oxygen species, respiratory chain f Abstract Complex II is the only membrane-bound component of the Krebs cycle and in addition functions as a member of the electron transport chain in mitochondria and in many bacteria. A recent X-ray structural solution of members of the complex II family of proteins has provided important insights into their function. One feature of the complex II structures is a linear electron transport chain that extends from the flavin and iron-sulfur redox cofactors in the membrane extrinsic domain to the quinone and b heme cofactors in the membrane domain. Exciting recent developments in relation to disease in humans and the formation of reactive oxygen species by complex II point to its overall importance in cellular physiology. CONTENTS OVERVIEW OF MEMBRANE-BOUND RESPIRATORY CHAIN .......... 78 COMPLEX II .......................................... 80 OVERALL STRUCTURE OF COMPLEX II ....................... 83 FLAVOPROTEIN SUBUNIT AND FORMATION OF THE COVALENT FAD LINKAGE............................................ 87 CATALYSIS IN COMPLEX II................................ 91 IRON-SULFUR CLUSTERS OF COMPLEX II AND THE ELECTRON TRANS- FER PATHWAY........................................ 94 MEMBRANE DOMAIN OF COMPLEX II ........................ 95 ROLE OF THE b HEME IN COMPLEX II ........................ 99 RELATION OF COMPLEX II TO DISEASE AND REACTIVE OXYGEN SPECIES ............................................100 CONCLUDING REMARKS .................................104 *The U.S.
    [Show full text]
  • Human Mitochondrial Pathologies of the Respiratory Chain and ATP Synthase: Contributions from Studies of Saccharomyces Cerevisiae
    life Review Human Mitochondrial Pathologies of the Respiratory Chain and ATP Synthase: Contributions from Studies of Saccharomyces cerevisiae Leticia V. R. Franco 1,2,* , Luca Bremner 1 and Mario H. Barros 2 1 Department of Biological Sciences, Columbia University, New York, NY 10027, USA; [email protected] 2 Department of Microbiology,Institute of Biomedical Sciences, Universidade de Sao Paulo, Sao Paulo 05508-900, Brazil; [email protected] * Correspondence: [email protected] Received: 27 October 2020; Accepted: 19 November 2020; Published: 23 November 2020 Abstract: The ease with which the unicellular yeast Saccharomyces cerevisiae can be manipulated genetically and biochemically has established this organism as a good model for the study of human mitochondrial diseases. The combined use of biochemical and molecular genetic tools has been instrumental in elucidating the functions of numerous yeast nuclear gene products with human homologs that affect a large number of metabolic and biological processes, including those housed in mitochondria. These include structural and catalytic subunits of enzymes and protein factors that impinge on the biogenesis of the respiratory chain. This article will review what is currently known about the genetics and clinical phenotypes of mitochondrial diseases of the respiratory chain and ATP synthase, with special emphasis on the contribution of information gained from pet mutants with mutations in nuclear genes that impair mitochondrial respiration. Our intent is to provide the yeast mitochondrial specialist with basic knowledge of human mitochondrial pathologies and the human specialist with information on how genes that directly and indirectly affect respiration were identified and characterized in yeast. Keywords: mitochondrial diseases; respiratory chain; yeast; Saccharomyces cerevisiae; pet mutants 1.
    [Show full text]
  • The Impact of CO2/HCO3 Availability on Anaplerotic Flux in PDHC
    bioRxiv preprint doi: https://doi.org/10.1101/663856; this version posted June 9, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. - 1 The impact of CO2/HCO3 availability on anaplerotic flux in PDHC- 2 deficient Corynebacterium glutamicum strains 3 a a a a b# 4 Aileen Krüger , Johanna Wiechert , Cornelia Gätgens , Tino Polen , Regina Mahr 5 and Julia Frunzkea# 6 7 a Institut für Bio- und Geowissenschaften, IBG-1: Biotechnology, Forschungszentrum 8 Jülich, 52425 Jülich, Germany 9 b SenseUp GmbH, c/o Campus Forschungszentrum Jülich, 52425 Jülich, Germany 10 - 11 Running Head: Influence of CO2/HCO3 on anaplerotic flux 12 13 A.K. and J.W. contributed equally to this work. 14 15 #Address correspondence to Julia Frunzke, [email protected], and Regina 16 Mahr, [email protected]. 17 1 bioRxiv preprint doi: https://doi.org/10.1101/663856; this version posted June 9, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 18 Abstract 19 The pyruvate dehydrogenase complex (PDHC) catalyzes the oxidative 20 decarboxylation of pyruvate yielding acetyl-CoA and CO2. The PDHC-deficient 21 Corynebacterium glutamicum strain ΔaceE is therefore lacking an important 22 decarboxylation step in central metabolism. Additional inactivation of pyc, encoding 23 pyruvate carboxylase, resulted in a >15 hour lag phase in the presence of glucose, 24 while no growth defect was observed on gluconeogenetic substrates like acetate.
    [Show full text]
  • Coupling and Pathway Control of Coenzyme Q Redox State
    MitoFit Preprints 2021.2.v4 doi:10.26124/mitofit:2021-0002.v4 Open Access Technical Communication Coupling and pathway control Cite Komlódi T, Cardoso LHD, of coenzyme Q redox state and Doerrier C, Moore AL, Rich PR, Gnaiger E (2021) respiration in isolated Coupling and pathway control of coenzyme Q redox mitochondria state and respiration in isolated mitochondria. 1 1 MitoFit Preprints 2021.2.v4. Timea Komlódi , Luiza HD Cardoso , doi:10.26124/mitofit:2021- Carolina Doerrier1, Anthony L Moore2, 0002.v4 Peter R Rich3, Erich Gnaiger1* Author contributions 1 Oroboros Instruments, Innsbruck, Austria; EG and TK collaborated 2 Biochemistry and Medicine, School of Life Sciences, closely with WGT in the Univ Sussex, Falmer, Brighton, UK; development of the Q- 3 Module, with guidance from Dept Structural and Molecular Biol, Univ College ALM and PRR. TK and LHDC London, London, UK designed, carried out and * Corresponding author: [email protected] analyzed the experiments. CD contributed to SUIT protocol development. TK, LHDC and Abstract EG co-wrote the manuscript. All authors commented on Redox states of the mitochondrial coenzyme Q and approved the pool, which reacts with the electron transfer system, manuscript. reflect the balance between (1) reducing capacities of Conflicts of interest electron flow from fuel substrates converging at the EG is founder and CEO of Q-junction, (2) oxidative capacities downstream of Q Oroboros Instruments, to O2, and (3) the load on the OXPHOS system utilizing Innsbruck, Austria. or dissipating the protonmotive
    [Show full text]
  • Coupling and Pathway Control of Coenzyme Q Redox State
    MitoFit Preprints 2021.2 doi:10.26124/mitofit:2021-0002 Open Access 1 Technical Communication 2 Coupling and pathway control of coenzyme Q 3 redox state and respiration in isolated 4 mitochondria 5 Timea Komlodi, Luiza HD Cardoso, Carolina Doerrier, Erich Gnaiger* 6 Oroboros Instruments, Innsbruck, Austria 7 8 * Corresponding author: [email protected] 9 10 Posted Online 2021-02-18 11 12 Abstract 13 Redox states of mitochondrial coenzyme Q (mtCoQ or Q) reflect the balance 14 between (1) reducing capacities of electron flow from fuel substrates converging at 15 the Q-junction, (2) oxidative capacities downstream of Q to oxygen, and (3) the load 16 on the OXPHOS system utilizing or dissipating the protonmotive force. A three- 17 electrode sensor (Rich 1988; Moore et al 1988) was implemented into the NextGen- 18 O2k to monitor the Q redox state continuously and simultaneously with oxygen 19 consumption. The Q-Module was optimized for high signal-to-noise ratio and 20 minimum oxygen diffusion. CoQ2 is added as a redox probe equilibrating with Q at 21 Complexes CI, CII and CIII and the detecting electrode. Q-sensors are poised with 22 the CoQ2 redox peak potentials determined by cyclic voltammetry, which provides 23 quality control of the Q-sensor and reveals chemical interferences. 24 25 The Q redox state and oxygen consumption were measured simultaneously in 26 isolated mitochondria. A coupling-control protocol was applied to analyze LEAK, 27 OXPHOS, and electron transfer capacities (L, P, and E, respectively) in the succinate- 28 pathway. In a second pathway-control protocol, NADH- and succinate-linked pathways 29 (N and S) converge at the Q-junction.
    [Show full text]
  • Mitochondrial Structure and Bioenergetics in Normal and Disease Conditions
    International Journal of Molecular Sciences Review Mitochondrial Structure and Bioenergetics in Normal and Disease Conditions Margherita Protasoni 1 and Massimo Zeviani 1,2,* 1 Mitochondrial Biology Unit, The MRC and University of Cambridge, Cambridge CB2 0XY, UK; [email protected] 2 Department of Neurosciences, University of Padova, 35128 Padova, Italy * Correspondence: [email protected] Abstract: Mitochondria are ubiquitous intracellular organelles found in almost all eukaryotes and involved in various aspects of cellular life, with a primary role in energy production. The interest in this organelle has grown stronger with the discovery of their link to various pathologies, including cancer, aging and neurodegenerative diseases. Indeed, dysfunctional mitochondria cannot provide the required energy to tissues with a high-energy demand, such as heart, brain and muscles, leading to a large spectrum of clinical phenotypes. Mitochondrial defects are at the origin of a group of clinically heterogeneous pathologies, called mitochondrial diseases, with an incidence of 1 in 5000 live births. Primary mitochondrial diseases are associated with genetic mutations both in nuclear and mitochondrial DNA (mtDNA), affecting genes involved in every aspect of the organelle function. As a consequence, it is difficult to find a common cause for mitochondrial diseases and, subsequently, to offer a precise clinical definition of the pathology. Moreover, the complexity of this condition makes it challenging to identify possible therapies or drug targets. Keywords: ATP production; biogenesis of the respiratory chain; mitochondrial disease; mi-tochondrial electrochemical gradient; mitochondrial potential; mitochondrial proton pumping; mitochondrial respiratory chain; oxidative phosphorylation; respiratory complex; respiratory supercomplex Citation: Protasoni, M.; Zeviani, M.
    [Show full text]
  • Respiratory Chain Activity and Protein Expression in Skeletal Muscles from High and Low Feed Efficient Swine Within the Same Genetic Line
    University of Arkansas, Fayetteville ScholarWorks@UARK Theses and Dissertations 7-2020 Respiratory Chain Activity and Protein Expression in Skeletal Muscles from High and Low Feed Efficient Swine within the Same Genetic Line Earsie L. Alcorn de Feliz University of Arkansas, Fayetteville Follow this and additional works at: https://scholarworks.uark.edu/etd Part of the Animal Experimentation and Research Commons, Cellular and Molecular Physiology Commons, Food Science Commons, and the Meat Science Commons Citation Alcorn de Feliz, E. L. (2020). Respiratory Chain Activity and Protein Expression in Skeletal Muscles from High and Low Feed Efficient Swine within the Same Genetic Line. Theses and Dissertations Retrieved from https://scholarworks.uark.edu/etd/3802 This Dissertation is brought to you for free and open access by ScholarWorks@UARK. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of ScholarWorks@UARK. For more information, please contact [email protected]. Respiratory Chain Activity and Protein Expression in Skeletal Muscles from High and Low Feed Efficient Swine within the Same Genetic Line A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Food Science by Earsie L. Alcorn de Feliz Union University Bachelor of Science in Biology, 1975 University of Arkansas Master of Science in Natural Science, 1977 July 2020 University of Arkansas This dissertation is approved for recommendation to the graduate council. Charles V. Maxwell, Ph.D. Dissertation Director Navam Hettiarachchy, Ph. D. Gisela F. Erf, Ph.D. Committee Member Committee Member Zelpha B. Johnson, Ph.D. Luke Howard, Ph.D. Committee Member Committee Member ABSTRACT Current as well as past studies have provided evidence for a connection between macroeconomic/agricultural and cellular/biochemical basis of feed efficiency (FE; gain:feed) in farm animal production.
    [Show full text]
  • Type of the Paper (Article
    Supplementary Materials Table S1. Details of proteins reported in Figure 2 panel A. List of proteins up- and down-regulated in samples with medium progressive motility with respect to high progressive motility group identified by LC-MS/MS analysis. Results are expressed as ratios of protein levels of asthenozoospermics to normozoospermics. UniProt Gene Protein Name Medium/High ID Name P20273 CD22 B-cell receptor CD22 0.14 Q8IYQ7 THNS1 Threonine synthase-like 1 0.19 Q99523 SORT Sortilin 0.22 P82933 RT09 28S ribosomal protein S9 0.26 Q14156 EFR3A Protein EFR3 homolog A 0.27 Q9BWH2 FUND2 FUN14 domain-containing protein 2 0.28 Q5RI15 COX20 Cytochrome c oxidase assembly protein COX20 0.31 P53597 SUCA Succinate‐CoA ligase 0.31 Q9BTX1 NDC1 Nucleoporin NDC1 0.33 P15529 MCP Membrane cofactor protein (TLX) 0.35 Q8TEM1 PO210 Nuclear pore membrane glycoprotein 210 0.36 Q8NBX0 SCPDL Saccharopine dehydrogenase-like oxidoreductase 0.37 Q9P2R7 SUCB1 Succinate‐CoA ligase [ADP-forming] subunit beta 0.37 Q99661 KIF2C Kinesin-like protein KIF2C 0.38 P02750 A2GL Leucine-rich alpha-2-glycoprotein 0.39 Q6UN15 FIP1 Pre-mRNA 3'-end-processing factor FIP1 0.40 P11310 ACADM Medium-chain specific acyl-CoA dehydrogenase 0.41 Q99798 ACON Aconitate hydratase 0.42 P55809 SCOT1 Succinyl-CoA:3-ketoacid coenzyme A transferase 1 0.43 P08574 CY1 Cytochrome c1 0.44 O14672 ADA10 Disintegrin and metalloproteinase domain-containing protein 10 0.46 Q13438 OS9 Protein OS-9 0.46 Q16698 DECR 2,4-dienoyl-CoA reductase 0.48 Q92820 GGH Gamma-glutamyl hydrolase 0.48 O75521 ECI2 Enoyl-CoA
    [Show full text]
  • Crosstalk Between Cytochrome Bc1 Complex and Mitochondrial Matrix Enzymes
    CROSSTALK BETWEEN CYTOCHROME BC1 COMPLEX AND MITOCHONDRIAL MATRIX ENZYMES By QIYU WANG Bachelor of Science in Cell and Molecular Biology University of Science and Technology of China Hefei, China 2004 Master of Science in Biochemistry University of Science and Technology of China Hefei, China 2007 Submitted to the Faculty of the Graduate College of the Oklahoma State University in partial fulfillment of the requirements for the Degree of DOCTOR OF PHILOSOPHY December, 2011 CROSSTALK BETWEEN CYTOCHROME BC1 COMPLEX AND MITOCHONDRIAL MATRIX ENZYMES Dissertation Approved: Dr. Chang-An Yu Dissertation Adviser Dr. Linda Yu Dr. José Luis Soulages Dr. Robert L. Burnap Outside Committee Member Dr. Sheryl A. Tucker Dean of the Graduate College ii TABLE OF CONTENTS Chapter Page I. INTRODUCTION .................................................................................................. 1 1.1 Introduction of cytochrome bc1 complex ..................................................... 2 1.1.1 Electron transfer chain............................................................................. 2 1.1.2 Complex III ............................................................................................. 5 1.1.3 Q-cycle mechanism ............................................................................... 12 1.1.4 Inhibitors of cytochrome bc1 ................................................................. 15 1.1.5 Other functions of cytochrome bc1 complex .......................................... 18 1.1.6 Interaction between matrix proteins
    [Show full text]
  • Structure, Function, and Assembly of Heme Centers in Mitochondrial Respiratory Complexes Hyung J
    University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Biochemistry -- Faculty Publications Biochemistry, Department of 2012 Structure, function, and assembly of heme centers in mitochondrial respiratory complexes Hyung J. Kim University of Utah Health Sciences Center Oleh Khalimonchuk University of Nebraska-Lincoln, [email protected] Pamela M. Smith University of Utah Health Sciences Center Dennis R. Winge University of Utah Health Sciences Center Follow this and additional works at: http://digitalcommons.unl.edu/biochemfacpub Part of the Biochemistry Commons, Biotechnology Commons, and the Other Biochemistry, Biophysics, and Structural Biology Commons Kim, Hyung J.; Khalimonchuk, Oleh; Smith, Pamela M.; and Winge, Dennis R., "Structure, function, and assembly of heme centers in mitochondrial respiratory complexes" (2012). Biochemistry -- Faculty Publications. 286. http://digitalcommons.unl.edu/biochemfacpub/286 This Article is brought to you for free and open access by the Biochemistry, Department of at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Biochemistry -- Faculty Publications by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. NIH Public Access Author Manuscript Biochim Biophys Acta. Author manuscript; available in PMC 2013 September 01. NIH-PA Author ManuscriptPublished NIH-PA Author Manuscript in final edited NIH-PA Author Manuscript form as: Biochim Biophys Acta. 2012 September ; 1823(9): 1604–1616. doi:10.1016/j.bbamcr.2012.04.008. Structure, function, and assembly of heme centers in mitochondrial respiratory complexes Hyung J. Kim, Oleh Khalimonchuk1, Pamela M. Smith, and Dennis R. Winge* University of Utah Health Sciences Center, Departments of Medicine and Biochemistry, Salt Lake City, UT 84132, USA Abstract The sequential flow of electrons in the respiratory chain, from a low reduction potential substrate to O2, is mediated by protein-bound redox cofactors.
    [Show full text]