Abstract
Cytochrome c oxidase (COX), which is located in the inner membrane of mitochondria, is a key constituent of the electron transport chain that catalyzes the reduction of oxygen. The Pacific whiteleg shrimp Litopenaeus vannamei is constantly exposed to hypoxic conditions, which affects both the central metabolism and the mitochondrial function. The purpose of this study was to isolate shrimp mitochondria, identify the COX complex and to evaluate the effect of hypoxia on the shrimp mitochondrial function and in the COX activity. A 190 kDa protein was identified as COX by immunodetection techniques. The effect of hypoxia was confirmed by an increase in the shrimp plasma L-lactate concentration. COX activity, mitochondrial oxygen uptake and protein content were reduced under hypoxic conditions, and gradually restored as hypoxia continued, this suggests an adaptive mitochondrial response and a highly effective COX enzyme. Both mitochondrial oxygen uptake and COX activity were completely inhibited by KCN and sodium azide, suggesting that COX is the unique oxidase in L. vannamei mitochondria.
Similar content being viewed by others
References
Abele D, Philip E, Gonzalez PM, Puntarulo S (2007) Marine invertebrate mitochondria and oxidative stress. Front Biosci 12:933–946
Buckler KJ, Turner PJ (2013) Oxygen sensitivity of mitochondrial function in rat arterial chemoreceptor cells. J Physiol 591(14):3549–3563
Chen J, Gao Y, Liao W, Huang J, Gao W (2012) Hypoxia affects mitochondrial protein expression in rat skeletal muscle. OMICS J Integr Biol 16(3):98–104
Costa-de-Oliveira S, Sampaio-Marques B, Barbosa M, Ricardo E, Pina-Vaz C, Ludovico P, Rodrigues AG (2012) An alternative respiratory pathway on Candida krusei: implications on susceptibility. Profile and oxidative stress. FEMS Yeast Res 20:287–294
Dabos KJ, Whalen HR, Newsome PN, Parkinson JA, Henderson NC, Sadler IH, Hayes PC, Plevris JN (2011) Impaired gluconeogenesis in a porcine model of paracetamol induced acute liver failure. World J Gastroenterol 17(11):1457–1461
Dassa EP, Dufour E, Gonçalves S, Paupe V, Hakkaart GA, Jacobs HT, Rustin P (2009) Expression of the alternative oxidase complements cytochrome c oxidase deficiency in human cells. EMBO Mol Med 1(1):30–36
Dudkina NV, Sunderhaus S, Boekema EJ, Braun HP (2008) The higher level of organization of the oxidative phosphorylation system: mitochondrial supercomplexes. J Bioenerg Biomembr 40(5):419–424
Fontanesi F, Soto IC, Barrientos A (2008) Cytochrome c oxidase biogenesis: new levels of regulation. Int Union Biochem Mol Biol Life 60(9):557–568
Fukuda R, Zhang H, Kim JW, Shimoda L, Dang CV, Semenza GL (2007) HIF-1 regulates cytochrome oxidase subunits to optimize efficiency of respiration in hypoxic cells. Cell 129:111–122
Grossman LI, Lomax MI (1997) Nuclear genes for cytochrome c oxidase. Biochim Biophys Acta 1352:171–192
Guerrero-Castillo S, Vazquez-Acevedo M, Gonzalez-Halphen D, Uribe-Carvajal S (2009) In Yarrowia lipolytica mitochondria, the alternative NADH dehydrogenase interacts specifically with the cytochrome complexes of the classic respiratory pathway. Biochim Biophys Acta 1787:75–85
Hildebrandt TM, Grieshaber MK (2008) Redox regulation of mitochondrial sulfide oxidation in the lugworm, Arenicola marina. J Exp Biol 211:2617–2623
Hochachka PW, Somero GN (2002) Influence of oxygen availability. In: Hochachka PW, Somero GN (eds) Biochemical adaptation. Oxford University Press, New York, pp 107–157
Hofmann GE, Hand SC (1990) Arrest of cytochrome-c oxidase synthesis coordinated with catabolic arrest in dormant Artemia embryos. Am J Physiol 258(5):R1184–R1191
Hüttemann M, Helling S, Sanderson TH, Sinkler C, Samavati L, Mahapatra G, Varughese A, Lu G, Liu J, Ramzan R, Vogt S, Grossman LI, Doan JW, Marcus K, Lee I (2012) Regulation of mitochondrial respiration and apoptosis through cell signalling: cytochrome c oxidase and cytochrome c in ischemia/reperfusion injury and inflammation. Biochim Biophys Acta 1817:598–609
Jimenez-Gutierrez LR, Hernandez-Lopez J, Islas-Osuna MA, Muhlia-Almazan A (2013) Three nucleus-encoded subunits of mitochondrial cytochrome c oxidase of the whiteleg shrimp Litopenaeus vannamei: cDNA characterization, phylogeny and mRNA expression during hypoxia and reoxygenation. Comp Biochem Physiol 166B(1):30–39
Keller M, Sommer AM, Pörtner KO, Abele D (2004) Seasonality of energetic functioning and production of reactive oxygen species by lugworm (Arenicola marina) mitochondria exposed to acute temperature changes. J Exp Biol 207(14):2529–2538
Liu JZ, Gao WX, Cai MC, Cao LF, Sun BY (2002) Effects of ATP concentration and hypoxic exposure on RNA and protein synthesis activity in isolated mitochondria from rat brain. Sheng Li Xue Bao 54(6):485–489
Martinez-Cruz O, Calderon de la Barca AM, Uribe-Carvajal S, Muhlia-Almazan A (2012) The function of mitochondrial FOF1 ATP-synthase from the whiteleg shrimp Litopenaeus vannamei muscle during hypoxia. Comp Biochem Physiol 162B:107–112
Mayevsky A, Rogatsky GG (2007) Mitochondrial function in vivo evaluated by NADH fluorescence: from animal models to human studies. Am J Physiol 292:C615–C640
McDonald AE, Vanlerberghe GC, Staples JF (2009) Alternative oxidase in animals: unique characteristics and taxonomic distribution. J Exp Biol 212(16):2627–2634
Ongwijitwat S, Wong-Riley MTT (2005) Is nuclear respiratory factor 2 a master transcriptional coordinator for all ten-nuclear encoded cytochrome c oxidase subunits in neurons? Gene 360:65–77
Paez-Osuna F, Gracia A, Flores-Verdugo F, Lyle-Fritch LP, Alonso-Rodrıguez R, Roque A, Ruiz-Fernandez AC (2003) Shrimp aquaculture development and the environment in the Gulf of California ecoregion. Mar Pollut Bull 46:806–815
Paital B, Chainy GB (2012) Effects of salinity on O2 consumption, ROS generation and oxidative stress status of gill mitochondria of the mud crab Scylla serrata. Comp Biochem Physiol 155C(2):228–237
Papandreou I, Cairns RA, Fontana L, Lim AL, Denko NC (2006) HIF-1 mediates adaptation to hypoxia by actively down-regulating mitochondrial oxygen consumption. Cell Metab 3(3):187–197
Philip E, Abele D (2007) Marine invertebrate mitochondria: respiratory balance in extreme environments. Comp Biochem Physiol 148A:S91–S94
Pierron D, Wildman DE, Hüttemann M, Markondapatnaikuni GC, Aras S, Grossman LI (2012) Cytochrome c oxidase: evolution of control via nuclear subunit addition. Biochim Biophys Acta 1817(4):590–597
Prabu SK, Anandatheerthavarada HK, Raza H, Srinivasan S, Spear JF, Avadhani NG (2006) Protein kinase A-mediated phosphorylation modulates cytochrome c oxidase function and augments hypoxia and myocardial ischemia-related injury. J Biol Chem 281(4):2061–2070
Racotta I, Palacios SE, Mendez L (2002) Metabolic responses to short and long term exposure to hypoxia in white shrimp (Penaeus vannamei). Mar Freshw Behav Physiol 35:269–275
Re AD, Diaz F (2011) Effect of different oxygen concentrations on physiological energetics of blue shrimp, Litopenaeus stylirostris (Stimpson). Open Zool J 4:1–8
Schägger H, von Jagow G (1995) Native electrophoresis for isolation of mitocondrial oxidative phosphorylation protein complexes. Methods Enzymol 260:190–203
Vargas-Albores F, Guzmán MA, Ochoa JL (1993) An anticoagulant solution for haemolymph collection and prophenoloxidase studies of penaeus shrimp. Comp Biochem Physiol 106A(2):299–303
Vijayasarathy C, Damle S, Prabu S, Otto C, Avadhani N (2003) Adaptive changes in the expression of nuclear and mitochondrial encoded subunits of cytochrome c oxidase and the catalytic activity during hypoxia. Eur J Biochem 270:871–879
Wittig I, Karas M, Schägger H (2007) High resolution clear native electrophoresis for in-gel functional assays and fluorescence studies of membrane protein complexes. Mol Cell Proteomics 6:1215–1225
Yang J, Zhu J, Xu WH (2010) Differential expression, phosphorylation of COX subunit 1 and COX activity during diapause phase in the cotton bollworm, Helicoverpa armigera. J Insect Physiol 56(12):1992–1998
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Jimenez-Gutierrez, L.R., Uribe-Carvajal, S., Sanchez-Paz, A. et al. The cytochrome c oxidase and its mitochondrial function in the whiteleg shrimp Litopenaeus vannamei during hypoxia. J Bioenerg Biomembr 46, 189–196 (2014). https://doi.org/10.1007/s10863-013-9537-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10863-013-9537-5