DOCSLIB.ORG

Myoglobin-Mediated Oxygen Delivery to Mitochondria of Isolated Cardiac Myocytes (Electron Transport/Heart Cells/Cytochrome Oxidase) BEATRICE A

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Myoglobin-Mediated Oxygen Delivery to Mitochondria of Isolated Cardiac Myocytes (Electron Transport/Heart Cells/Cytochrome Oxidase) BEATRICE A Proc. Nati. Acad. Sci. USA Vol. 84, pp. 7503-7507, November 1987 Biochemistry Myoglobin-mediated oxygen delivery to mitochondria of isolated cardiac myocytes (electron transport/heart cells/cytochrome oxidase) BEATRICE A. WITTENBERG* AND JONATHAN B. WITTENBERG Department of Physiology and Biophysics, Albert Einstein College of Medicine, Bronx, NY 10461 Communicated by Berta Scharrer, July 20, 1987 (receivedfor review May 5, 1987) ABSTRACT Myoglobin-mediated oxygen delivery to in- Cytochrome oxidase, half-oxidized when ambient oxygen tracellular mitochondria is demonstrated in cardiac myocytes partial pressure (Po2) is 0.07 torr (1 torr = 133 Pa) (16), in the isolated from the hearts of mature rats. Myocytes are held at circumstance described here experiences oxygen pressures high ambient oxygen pressure, 40-340 torr (5-45 kPa); 20- to 200-fold the pressure required to maintain the normal, sarcoplasmic myoglobin is fully oxygenated. In this condition largely oxidized, state seen in resting myocytes (16). Carbon oxygen availability does not limit respiratory rate; myoglobin- monoxide in this circumstance blocks oxygenation of facilitated diffusion contributes no additional oxygen flux and, sarcoplasmic myoglobin selectively without perturbing the since oxygen consumption is measured in steady states, the optical spectrum of intracellular cytochrome oxidase. We storage function of myoglobin vanishes. Carbon monoxide, conclude that cardiac mitochondria accept two additive introduced stepwise, displaces oxygen from intracellular simultaneous flows of oxygen: the well-known flow of dis- oxymyoglobin without altering the optical spectrum of the solved oxygen to cytochrome oxidase and a flow of largely oxidized intracellular mitochondria. A large part, myoglobin-bound oxygen to a mitochondrial terminus. The about one-third, of the steady-state oxygen uptake is abolished myoglobin-mediated oxygen flow supports ATP generation by carbon monoxide blockade of myoglobin oxygenation. The in the physiological range of oxygen pressure. myoglobin-dependent component of the oxygen uptake de- creases linearly with decreasing fraction of intracellular MATERIALS AND METHODS oxymyoglobin, with a slope near unity. Studies using inhibitors of mitochondrial electron transport indicate- that myoglobin- Isolated Cardiac Myocytes. These were prepared from the delivered oxygen uptake depends on electron flow through the hearts of mature rats by enzymatic digestion, purified on mitochondrial electron transport chain. We conclude that Percoll (Pharmacia) density gradients, and suspended to a cardiac mitochondria accept two additive simultaneous flows of final density of 0.5-1.0 x 106 cells per ml (19, 20). oxygen: a flow of dissolved oxygen to cytochrome oxidase and Mitochondria. Mitochondria were prepared by the method a flow of myoglobin-bound oxygen to a mitochondrial termi- of Palmer et al. (21) from adult rat hearts that had been nus. Myoglobin-mediated oxygen delivery supports ATP gen- perfused with balanced saline solution to remove erythro- eration by heart cells at physiological ambient oxygen pressure. cytes. Gas Partial Pressures and Oxygen Uptake. All measure- Myoglobin is an oxygen-binding monomeric hemeprotein ments were made in steady states (7, 16) of constant oxygen found in the cytosol of cardiac myocytes and of those pressure and oxygen uptake. The measuring chamber had vertebrate muscle fibers that do sustained work (1). The both liquid and gas phases (16, 22). The gas phase composi- myoglobin content of muscles increases with exercise (1-3) tion was set by a mass flow controller (Tylan, Torrance, CA). and is proportional to the cytochrome oxidase content (2, 4, Since carbon monoxide is not consumed, solution Pco is 5). Myoglobin, by facilitating oxygen diffusion, maintains an known from the composition of the gas phase. Solution ample free oxygen concentration at the muscle mitochon- oxygen pressure was monitored by a sensitive polarographic drion (2, 4, 6). Blockade of myoglobin function decreases oxygen electrode and is the balance of oxygen entering and oxygen uptake (7, 8), work output (8, 9), and ATP generation oxygen consumed by the myocytes. At constant temperature (10) of cardiac and skeletal muscle. and stirring rate, oxygen uptake is known from the difference Facilitation of oxygen diffusion through the sarcoplasm of between solution Po2 in the absence of myocytes (equal to the myocyte has often been taken as the sole function of gas-phase Po2) and the actual solution Po2 in the presence of myoglobin in muscles in the steady state. Here we show an myocytes, using a mass transfer coefficient that is determined additional function: myoglobin-mediated oxygen delivery to daily (16, 22, 23). The temperature was 30'C in all experi- mitochondria. This is most clearly demonstrated in isolated ments. cardiac myocytes flooded with superabundant oxygen (11, Optical Spectra. The chamber was held in a thermostatted 12). In this circumstance, the maximum diffusive flow of block placed in the sample light beam of a Cary model 17 dissolved oxygen across the sarcoplasm exceeds the mito- recording spectrophotometer (Varian) equipped with a Cary chondrial oxygen demand at least 100-fold (13, 14); sarco- scattered transmission accessory. Data were acquired digi- plasmic myoglobin is essentially fully oxygenated, and facil- tally from 650 to 350 nm, and difference spectra were itated diffusion contributes no additional oxygen flux (2, 4, 6). constructed by subtraction, using an Aviv data acquisition Since oxygen uptake is in steady states, the storage function system (Aviv Laboratories, Lakewood, NJ). The spectral of myoglobin vanishes (7). Intracellular gradients of oxygen contribution of myoglobin dominates the myocyte optical pressure, always shallow (15-18), now are small relative to spectrum (16). Under the conditions of our experiments, ambient oxygen pressure. Abbreviations: CCCP, carbonyl cyanide m-chlorophenylhydrazone; MbO2, oxymyoglobin; MbCO, carbon monoxide myoglobin; Po2, The publication costs of this article were defrayed in part by page charge partial pressure of oxygen; Pco, partial pressure of carbon monox- payment. This article must therefore be hereby marked "advertisement" ide. in accordance with 18 U.S.C. §1734 solely to indicate this fact. *To whom reprint requests should be addressed. 7503 Downloaded by guest on October 1, 2021 7504 Biochemistry: Wittenberg and Wittenberg Proc. Natl. Acad Sci. USA 84 (1987) intracellular myoglobin is essentially completely ligated; oxy- a and carbon monoxide myoglobin are the dominant species present. The contribution of myoglobin was calculated from w the spectral changes occurring at 409 and 424 nm, when By intracellular oxymyoglobin was converted fully to carbon 0u monoxide myoglobin, and was subtracted from myocyte spectra to obtain a residuum dominated by the spectral z contribution of the mitochondria. The fraction of carbon w monoxide myoglobin at any Pco was calculated from spectral x changes observed at 409 and 424 nm. Excellent isosbesticity 0 was maintained at 415 and 438 nm. Energetics of Cardiac Myocytes. Suspensions of myocytes were incubated in the measuring chamber for 60 min. Oxygen ATP, intracellular phosphocreatine, and b uptake, intracellular 0 1.0I accumulated lactate (final minus initial) were determined as 0 previously (16, 20). me - _ _ 0 U *- - - S~ RESULTS w A.~ I- 0.5 /U/ The partition of intracellular myoglobin between carbon a. ,0 monoxide and oxygen, M = ([MbCO] x Po2)/([MbO2] x z // Pco), was determined graphically as the slope of the line in 0/ Fig. 1. 'The partition coefficient, M = 20 at 30'C, is not largely / different from that of purified rat heart myoglobin, M = 29 at 0 00 I40 30'C, which we determined by conventional procedures (24, 0 10 20 30 40 25). The reasonable value of the slope, together with the Pco torr linearity of the relation found, indicates near equilibrium between sarcoplasmic myoglobin, oxygen, and carbon mon- FIG. 2. Steady-state oxygen uptake of suspensions of cardiac oxide. myocytes as a function of carbon monoxide partial pressure. Res- The effect of carbon monoxide on steady-state respiration piratory rate [14.4 ± 3.3 nmol of oxygen per min per ml per 106 cells of cardiac myocytes is presented in Fig. 2. Within each (mean ± SD; n = 29) in these experiments] is normalized, taking the experiment, the fraction of oxygen in the gas phase was held uninhibited rate in each experiment as unity. Data are from three experiments. m, 15% oxygen in the gas phase (solution P92 = 80-90 constant at 10-25% and the fraction of carbon monoxide was torr); e, 20% oxygen (solution Po2 = 100-120 torr); A, 25% oxygen increased stepwise. Steady-state solution oxygen pressure, (solution Po2 = 155-160 torr). (a) Pco = 0-730 torr. (b) The same steady-state oxygen uptake, and optical spectra were moni- data presented over a limited range; the fractional saturation of tored. The oxygen uptake of cardiac myocytes held at high sarcoplasmic myoglobin in one experiment is presented (o--- o; Po2 decreases initially as Pco is increased, stays nearly 20%o oxygen in the gas phase). The myoglobin-dependent component constant a in which is of the oxygen uptake is taken as the difference between the in prolonged plateau myoglobin fully uninhibited rate and the plateau value reached as sarcoplasmic saturated with carbon monoxide, and finally, above 95% myoglobin approaches saturation with carbon monoxide. carbon monoxide, in the gas phase, declines to zero as carbon
Load more

Recommended publications
  • SUPPLEMENTARY DATA Data Supplement To
    SUPPLEMENTARY DATA Data supplement to Perivascular adipose tissue controls insulin-stimulated perfusion, mitochondrial protein expression and glucose uptake in muscle through adipomuscular microvascular anastomoses Surname first author Turaihi Authors & Affiliations Turaihi, Alexander H MD1; Serné, Erik H, MD PhD2; Molthoff, Carla FM PhD3; Koning, Jasper J PhD4; Knol, Jaco PhD6; Niessen, Hans W MD PhD5; Goumans, Marie Jose TH PhD7; van Poelgeest, Erik M MD1; Yudkin, John S MD PhD8; Smulders, Yvo M MD PhD2; Connie R Jimenez, PhD6; van Hinsbergh, Victor WM PhD1; Eringa, Etto C PhD1 ©2020 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db18-1066/-/DC1 SUPPLEMENTARY DATA Western immunoblotting Skeletal muscle samples were lysed up in 1D‐sample buffer (10% glycerol, 62.5 mmol/L Tris (pH 6.8), 2% w/v LDS, 2% w/v DTT) and protein concentration was determined using Pierce 660‐nm protein assay (Thermo scientific, Waltham, MA USA 02 451; 22 660) according to the manufacturer's instructions. Heat shock protein 90 immunoblotting was performed by application of samples (5 µg protein) on 4‐15% Criterion TGX gels (Biorad, Veenendaal, the Netherlands, 5 671 084) and semi‐dry blotting onto PVDF membranes (GE Healthcare‐Fisher, RPN1416F), incubated overnight with rat monoclonal HSP90 antibody (1:1000 dilution) after blocking with 5% milk in TBS‐T (137 mM NaCl, 20 mmol/L Tris pH 7.0 and 0.1% (v/v) Tween [Sigma‐Aldrich, P7949]). After 2 hours incubation with anti-rat, horse radish peroxidase-coupled secondary antibody (Thermo Fisher 62-9520), the blot was stained using ECL‐prime (Fisher scientific, 10 308 449) and analysed on an AI‐600 imaging system (GE Healthcare, Life Sciences).
    [Show full text]
  • Table 1. Identified Proteins with Expression Significantly Altered in the Hippocampus of Rats of Exposed Group (Pb) Vs
    Table 1. Identified proteins with expression significantly altered in the hippocampus of rats of exposed group (Pb) vs. Control. Fold Change Accession Id a Protein Description Score Pb P35213 14-3-3 protein beta/alpha 85420 −0.835 P62260 14-3-3 protein epsilon 96570 −0.878 P68511 14-3-3 protein eta 85420 −0.844 P68255 14-3-3 protein theta 85420 −0.835 P63102 14-3-3 protein zeta/delta 105051 −0.803 P13233 2',3'-cyclic-nucleotide 3'-phosphodiesterase 151400 1.405 P68035 Actin, alpha cardiac muscle 1 442584 −0.942 P68136 Actin, alpha skeletal muscle 441060 −0.970 P62738 Actin, aortic smooth muscle 438270 −0.970 P60711 Actin, cytoplasmic 1 630104 −0.942 P63259 Actin, cytoplasmic 2 630104 −0.942 P63269 Actin, gamma-enteric smooth muscle 438270 −0.951 Q05962 ADP/ATP translocase 1 60100 −0.554 Q09073 ADP/ATP translocase 2 49102 −0.482 P84079 ADP-ribosylation factor 1 34675 −0.644 P84082 ADP-ribosylation factor 2 22412 −0.644 P61206 ADP-ribosylation factor 3 34675 −0.619 P61751 ADP-ribosylation factor 4 22412 −0.670 P84083 ADP-ribosylation factor 5 22412 −0.625 P04764 Alpha-enolase 46219 −0.951 P23565 Alpha-internexin 9478 1.062 P37377 Alpha-synuclein 89619 −0.771 P13221 Aspartate aminotransferase, cytoplasmic 23661 1.083 P00507 Aspartate aminotransferase, mitochondrial 46049 1.116 P10719 ATP synthase subunit beta, mitochondrial 232442 −0.835 P85969 Beta-soluble NSF attachment protein 9638 1.419 Q63754 Beta-synuclein 66842 −0.779 P11275 Calcium/calmodulin-dependent protein kinase type II subunit alpha 181954 1.105 P08413 Calcium/calmodulin-dependent protein kinase type II subunit beta 80840 1.127 P15791 Calcium/calmodulin-dependent protein kinase type II subunit delta 62682 1.105 Int.
    [Show full text]
  • Iron Deficiency in the Rat: Effects on Neutrophil Activation and Metabolism
    THEOPHYLLINE AND BRAIN 549 A dietary protein deficiency can affect the susceptibility to the metabolism in children with protein-calorie malnutrition. Am. J. Clin. Nutr., 28: 977 (1975). toxicity of drugs or other agents such as pesticides and herbicides 8. Nakamoto. T. and Miller. S. A.: Effect of vrotein-energy malnutrition on the (2). At the present time, there are no such studies relative to growth df mandible aid long bone in newborn miie and female rats. J. theophylline, although the usage of theophylline is now quite Nutr., 107: 983 (1977). common in the neonatal intensive care environment. Many 9. Nebron, R. M., Resnick, M. D., and Halstmm, W. J.: Developmental outcome of premature infants treated with theophylline. Dev. Pharmacol. Ther., 1: infants therein who are now receiving theophylline therapeuti- 274 ( 1980). cally are being dosed on a body weight basis without considera- 10. Newberne, P. M., Gross, R. L., and Roe, D. A,: Dmg, toxin, nutrient interac- tion of the nutritional status. Our data suggest that, in the animal tions. World Rev. Nutr. Diet., 29: 130 (1978). model, the administration of theophylline in the presence of a 1 I. Pastorova, B., Sova, O., and Burda, J.: Incorporation of I4C-thymidine into liver and brain DNA of protein-deficient rats. Physiol. Bohemoslov., 27: 69 compromised nutritional status may have effects not now appar- (1978). ent. We add our concern to that expressed by others (16) that 12. Prasad, A. S., Dumouchell, E., Kovich, D., and Oberleas, D.: A simple methylxanthine administration may have previously unsus- fluorometric method for the determination of RNA and DNA in tissue.
    [Show full text]
  • Chain of Human Neutrophil Cytochrome B CHARLES A
    Proc. Nati. Acad. Sci. USA Vol. 85, pp. 3319-3323, May 1988 Biochemistry Primary structure and unique expression of the 22-kilodalton light chain of human neutrophil cytochrome b CHARLES A. PARKOS*, MARY C. DINAUERt, LESLIE E. WALKER*, RODGER A. ALLEN*, ALGIRDAS J. JESAITIS*, AND STUART H. ORKINtt *Department of Immunology, Research Institute of the Scripps Clinic, La Jolla, CA 92037; tDivision of Hematology-Oncology, Children's Hospital, and Dana-Farber Cancer Institute, Department of Pediatrics, Harvard Medical School, Boston, MA 02115; and tHoward Hughes Medical Institute, Children's Hospital, Boston, MA 02115 Communicated by Harvey F. Lodish, January 14, 1988 ABSTRACT Cytochrome b comprising 91-kDa and 22- Cytochrome b purified from neutrophil membranes ap- kDa subunits is a critical component of the membrane-bound pears to be a heterodimer of a glycosylated 91-kDa heavy oxidase of phagocytes that generates superoxide. This impor- chain and a nonglycosylated 22-kDa light chain (10-12). The tant microbicidal system is impaired in inherited disorders 91-kDa subunit is encoded by a gene designated CGD, known as chronic granulomatous disease (CGD). Previously we residing at chromosomal position Xp2l, which originally was determined the sequence of the larger subunit from the cDNA identified on the basis of genetic linkage without reference to of the CGD gene, the X chromosome locus affected in "X- a specific protein product (8). Antisera generated to either a linked" CGD. To complete the primary structure of the synthetic peptide predicted from the cDNA or to a fusion cytochrome b and to assess expression of the smaller subunit, protein produced in E.
    [Show full text]
  • Table S1. Identified Proteins with Exclusive Expression in Cerebellum of Rats of Control, 10Mg F/L and 50Mg F/L Groups
    Table S1. Identified proteins with exclusive expression in cerebellum of rats of control, 10mg F/L and 50mg F/L groups. Accession PLGS Protein Name Group IDa Score Q3TXS7 26S proteasome non-ATPase regulatory subunit 1 435 Control Q9CQX8 28S ribosomal protein S36_ mitochondrial 197 Control P52760 2-iminobutanoate/2-iminopropanoate deaminase 315 Control Q60597 2-oxoglutarate dehydrogenase_ mitochondrial 67 Control P24815 3 beta-hydroxysteroid dehydrogenase/Delta 5-->4-isomerase type 1 84 Control Q99L13 3-hydroxyisobutyrate dehydrogenase_ mitochondrial 114 Control P61922 4-aminobutyrate aminotransferase_ mitochondrial 470 Control P10852 4F2 cell-surface antigen heavy chain 220 Control Q8K010 5-oxoprolinase 197 Control P47955 60S acidic ribosomal protein P1 190 Control P70266 6-phosphofructo-2-kinase/fructose-2_6-bisphosphatase 1 113 Control Q8QZT1 Acetyl-CoA acetyltransferase_ mitochondrial 402 Control Q9R0Y5 Adenylate kinase isoenzyme 1 623 Control Q80TS3 Adhesion G protein-coupled receptor L3 59 Control B7ZCC9 Adhesion G-protein coupled receptor G4 139 Control Q6P5E6 ADP-ribosylation factor-binding protein GGA2 45 Control E9Q394 A-kinase anchor protein 13 60 Control Q80Y20 Alkylated DNA repair protein alkB homolog 8 111 Control P07758 Alpha-1-antitrypsin 1-1 78 Control P22599 Alpha-1-antitrypsin 1-2 78 Control Q00896 Alpha-1-antitrypsin 1-3 78 Control Q00897 Alpha-1-antitrypsin 1-4 78 Control P57780 Alpha-actinin-4 58 Control Q9QYC0 Alpha-adducin 270 Control Q9DB05 Alpha-soluble NSF attachment protein 156 Control Q6PAM1 Alpha-taxilin 161
    [Show full text]
  • Liver Mitochondrial DNA Damage and Genetic Variability of Cytochrome B - a Key Component of the Respirasome - Drive the Severity of Fatty Liver Disease
    DR. SILVIA SOOKOIAN (Orcid ID : 0000-0001-5929-5470) Article type : Original This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differencesAccepted Article between this version and the Version of Record. Please cite this article as doi: 10.1111/JOIM.13147 This article is protected by copyright. All rights reserved Liver mitochondrial DNA damage and genetic variability of Cytochrome b - a key component of the respirasome - drive the severity of fatty liver disease Short title: NASH and respirasome Carlos J Pirola *, Martin Garaycoechea, Diego Flichman, Gustavo O Castaño and Silvia Sookoian *. 1 University of Buenos Aires, School of Medicine, Institute of Medical Research A Lanari, Ciudad Autónoma de Buenos Aires, Argentina (CJP, SS). 2 National Scientific and Technical Research Council (CONICET)−University of Buenos Aires, Institute of Medical Research (IDIM), Department of Molecular Genetics and Biology of Complex Diseases, Ciudad Autónoma de Buenos Aires, Argentina (CJP). 3 Department of Surgery, Hospital de Alta Complejidad en Red “El Cruce”, Florencio Varela, Buenos Aires, Argentina (MG). 4 Department of Virology, School of Pharmacy and Biochemistry, University of Buenos Aires, Ciudad Autónoma de Buenos Aires, Argentina (DF). 5 Liver Unit, Medicine and Surgery Department, Hospital Abel Zubizarreta, Ciudad Autónoma de Buenos Aires, Argentina (GOC). 6 National Scientific and Technical Research Council (CONICET)−University of Buenos Aires, Institute of Medical Research (IDIM), Department of Clinical and Molecular Hepatology, Ciudad Autónoma de Buenos Aires, Argentina (SS). * Co-corresponding authorship Grant support: This study was partially supported by grants PICT 2015-0551 and PICT 2016- 0135 (Agencia Nacional de Promoción Científica y Tecnológica, FONCyT).
    [Show full text]
  • Prediction of the Amount of Secondary Structure in a Globular Protein from Its Aminoacid Compositionj (Helix/Pl-Sheet/Turns) W
    Proc. Nat. Acad. Sci. USA Vol. 70, No. 10, pp. 2809-2813, October 1973 Prediction of the Amount of Secondary Structure in a Globular Protein from Its Aminoacid Compositionj (helix/pl-sheet/turns) W. R. KRIGBAUM AND SARA PARKEY KNLTTTON Gross Chemical Laboratory, Duke University, Durham, North Carolina 27706 Communicated by Walter Gordy, June 22, 1973 ABSTRACT Multiple regression is used to obtain rela- so reference was made to the published crystal structures tionships for predicting the amount of secondary structure where ambiguities arose. Regions not included in the above in a protein molecule from a knowledge of its aminoacid composition. We tested these relations using 18 proteins of categories were termed coil. Assignments of secondary struc- known structure, but omitting the protein to be predicted. tural regions appear in Table 1. Since residues may be assipned Independent predictions were made for the two sub- to more than one category (e.g., the last residues of a helical chains of hemoglobin and insulin. The average errors for region and the first ones of a turn), then percentages do not these 20 chains or subchains are: helix ± 7.1%, p-sheet necessarily add up to 100% for any protein. ± 6.9%, turn ± 4.2%, and coil ± 5.7%. A second set of rela- tions yielding somewhat inferior predictions is given for We recognize at the outset that this data base contains the case in which Asp and Asn, and Glu and Gln, are not errors of at least two types. First, there are some remaining differentiated. Predictions are also listed for 15 proteins uncertainties in the primary structures.
    [Show full text]
  • Molecular Evolution of Cytochrome B in High- and Low-Altitude Deer Mice (Genus Peromyscus)
    University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Jay F. Storz Publications Papers in the Biological Sciences 2008 Molecular evolution of cytochrome b in high- and low-altitude deer mice (genus Peromyscus) E.J. Gering University of Texas, 1 University Station C0930, Austin, TX, [email protected] J. C. Opazo Instituto de Ecologia y Evolucion, Facultad de Ciencias, Universidad Austral de Chile, Casilla 567, Valdivia, Chile Jay F. Storz University of Nebraska - Lincoln, [email protected] Follow this and additional works at: https://digitalcommons.unl.edu/bioscistorz Part of the Genetics and Genomics Commons Gering, E.J.; Opazo, J. C.; and Storz, Jay F., "Molecular evolution of cytochrome b in high- and low-altitude deer mice (genus Peromyscus)" (2008). Jay F. Storz Publications. 23. https://digitalcommons.unl.edu/bioscistorz/23 This Article is brought to you for free and open access by the Papers in the Biological Sciences at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Jay F. Storz Publications by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. Published in Heredity (2008), doi: 10.1038/hdy.2008.124 Copyright © 2008 The Genetics Society; published by Macmillan/Nature Publishing Group. Used by permision. http://www.nature.com/hdy/index.html Submitted March 19, 2008; revised November 11, 2008; accepted November 21, 2008; published online December 24, 2008. Molecular evolution of cytochrome b in high- and low-altitude deer mice (genus Peromyscus) E. J. Gering,* J. C. Opazo,† and J. F. Storz School of Biological Sciences, University of Nebraska–Lincoln, Lincoln, NE, USA * Corresponding author — current address: Section of Integrative Biology, University of Texas, 1 University Station C0930, Austin, TX 78712, USA; email [email protected] † Current address: Instituto de Ecologia y Evolucion, Facultad de Ciencias, Universidad Austral de Chile, Casilla 567, Valdivia, Chile.
    [Show full text]
  • Molecular and Whole Animal Responses of Grass Shrimp, Palaemonetes Pugio, Exposed to Chronic Hypoxia ⁎ Marius Brouwer A, , Nancy J
    Journal of Experimental Marine Biology and Ecology 341 (2007) 16–31 www.elsevier.com/locate/jembe Molecular and whole animal responses of grass shrimp, Palaemonetes pugio, exposed to chronic hypoxia ⁎ Marius Brouwer a, , Nancy J. Brown-Peterson a, Patrick Larkin b, Vishal Patel c, Nancy Denslow c, Steve Manning a, Theodora Hoexum Brouwer a a Department of Coastal Sciences, The University of Southern Mississippi, 703 East Beach Dr., Ocean Springs, MS 39564, USA b EcoArray Inc., 12085 Research Dr., Alachua, Florida 32615, USA c Department of Physiological Sciences and Center for Environmental and Human Toxicology, University of Florida, PO Box 110885, Gainesville, FL 32611, USA Received 28 July 2006; received in revised form 15 September 2006; accepted 20 October 2006 Abstract Hypoxic conditions in estuaries are one of the major factors responsible for the declines in habitat quality. Previous studies examining effects of hypoxia on crustacea have focused on individual/population-level, physiological or molecular responses but have not considered more than one type of response in the same study. The objective of this study was to examine responses of grass shrimp, Palaemonetes pugio, to moderate (2.5 ppm DO) and severe (1.5 ppm DO) chronic hypoxia at both the molecular and organismal levels. At the molecular level we measured hypoxia-induced alterations in gene expression using custom cDNA macroarrays containing 78 clones from a hypoxia- responsive suppression subtractive hybridization cDNA library. Grass shrimp exposed to moderate hypoxia show minimal changes in gene expression. The response after short-term (3 d) exposure to severe hypoxia was up-regulation of genes involved in oxygen uptake/transport and energy production, such as hemocyanin and ATP synthases.
    [Show full text]
  • Roles for Enteric D-Type Cytochrome Oxidase in N2 Fixation and Microaerobiosis SUSAN HILL,'* SABINE VIOLLET,1 ANDREW T
    JOURNAL OF BACTERIOLOGY, Apr. 1990, p. 2071-2078 Vol. 172, No. 4 0021-9193/90/042071-08$02.00/0 Copyright C) 1990, American Society for Microbiology Roles for Enteric d-Type Cytochrome Oxidase in N2 Fixation and Microaerobiosis SUSAN HILL,'* SABINE VIOLLET,1 ANDREW T. SMITH,2t AND CHRISTOPHER ANTHONY2 Nitrogen Fixation Laboratory, AFRC Institute ofPlant Science Research, University of Sussex, Brighton BNJ 9RQ, United Kingdom,1 and Department ofBiochemistry, University of Southampton, Southampton S09 3TU, United Kingdom2 Received 26 September 1989/Accepted 3 January 1990 Escherichia coli strains that lacked the d-type cytochrome oxidase, the terminal oxidase with a high affinity for 02, grew anaerobically as well as the wild type did and were not impaired in the ability to evolve H2 from either glucose or formate. The anaerobic synthesis and activity of nitrogenase in transconjugants of these strains carrying Klebsiella pneumoniae nif genes were also normal. However, the behavior towards 02 of anaerobically grown bacteria lacking the d-type oxidase differed from that of the wild type in the following ways: the potential 02 uptake was lower, H2 evolution and nitrogenase activity supported by fermentation were more strongly inhibited by 02, and microaerobic 02-dependent nitrogenase activity in the absence of a fermentable carbon source did not occur. These results show that the d-type oxidase serves two functions in enteric bacteria-to conserve energy under microaerobic conditions and to protect anaerobic processes from inhibition by 02 A branched respiratory chain terminating in oxidases of activity (30 nM) is near the apparent Km of the purified differing affinities for 02 is common among procaryotes (1, d-type oxidase complex (20 nM 02) (20, 39a).
    [Show full text]
  • Novel Protein Pathways in Development and Progression of Pulmonary Sarcoidosis Maneesh Bhargava1*, K
    www.nature.com/scientificreports OPEN Novel protein pathways in development and progression of pulmonary sarcoidosis Maneesh Bhargava1*, K. J. Viken1, B. Barkes2, T. J. Grifn3, M. Gillespie2, P. D. Jagtap3, R. Sajulga3, E. J. Peterson4, H. E. Dincer1, L. Li2, C. I. Restrepo2, B. P. O’Connor5, T. E. Fingerlin5, D. M. Perlman1 & L. A. Maier2 Pulmonary involvement occurs in up to 95% of sarcoidosis cases. In this pilot study, we examine lung compartment-specifc protein expression to identify pathways linked to development and progression of pulmonary sarcoidosis. We characterized bronchoalveolar lavage (BAL) cells and fuid (BALF) proteins in recently diagnosed sarcoidosis cases. We identifed 4,306 proteins in BAL cells, of which 272 proteins were diferentially expressed in sarcoidosis compared to controls. These proteins map to novel pathways such as integrin-linked kinase and IL-8 signaling and previously implicated pathways in sarcoidosis, including phagosome maturation, clathrin-mediated endocytic signaling and redox balance. In the BALF, the diferentially expressed proteins map to several pathways identifed in the BAL cells. The diferentially expressed BALF proteins also map to aryl hydrocarbon signaling, communication between innate and adaptive immune response, integrin, PTEN and phospholipase C signaling, serotonin and tryptophan metabolism, autophagy, and B cell receptor signaling. Additional pathways that were diferent between progressive and non-progressive sarcoidosis in the BALF included CD28 signaling and PFKFB4 signaling. Our studies demonstrate the power of contemporary proteomics to reveal novel mechanisms operational in sarcoidosis. Application of our workfows in well-phenotyped large cohorts maybe benefcial to identify biomarkers for diagnosis and prognosis and therapeutically tenable molecular mechanisms.
    [Show full text]
  • RNA-Sequencing Indicates High Hemocyanin Expression As a Key Strategy for Cold Adaptation in the Antarctic Amphipod Eusirus Cf
    BIOCELL Tech Science Press 2021 RNA-sequencing indicates high hemocyanin expression as a key strategy for cold adaptation in the Antarctic amphipod Eusirus cf. giganteus clade g3 SAMUELE GRECO1,#;ELISA D’AGOSTINO2,#;CHIARA MANFRIN1;ANASTASIA SERENA GAETANO1;GAEL FURLANIS1; FRANCESCA CAPANNI1;GIANFRANCO SANTOVITO2;PAOLO EDOMI1;PIERO GIULIO GIULIANINI1;MARCO GERDOL1,* 1 Department of Life Sciences, University of Trieste, Trieste, 34127, Italy 2 Department of Biology, University of Padova, Padova, 35131, Italy Key words: Antarctica, Cold adaptation, Hemocyanin, Amphipod, Transcriptome Abstract: We here report the de novo transcriptome assembly and functional annotation of Eusirus cf. giganteus clade g3, providing the first database of expressed sequences from this giant Antarctic amphipod. RNA-sequencing, carried out on the whole body of a single juvenile individual likely undergoing molting, revealed the dominant expression of hemocyanins. The mRNAs encoding these oxygen-binding proteins cumulatively accounted for about 40% of the total transcriptional effort, highlighting the key biological importance of high hemocyanin production in this Antarctic amphipod species. We speculate that this observation may mirror a strategy previously described in Antarctic cephalopods, which compensates for the decreased ability to release oxygen to peripheral tissues at sub-zero temperatures by massively increasing total blood hemocyanin content compared with temperate species. These preliminary results will undoubtedly require confirmation through proteomic
    [Show full text]