Replacement of a cytosolic copper/zinc superoxide dismutase by a novel cytosolic manganese superoxide dismutase in crustaceans that use copper (haemocyanin) for oxygen transport

Brouwer, Marius; Brouwer, Thea Hoexum; Grater, Walter D.; Brown‐Peterson, Nancy J.

doi:10.1042/bj20030272

Cited by 101 publications

(79 citation statements)

References 43 publications

(67 reference statements)

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“…Ten genes (Table 1: genes 1 to 10), including heat shock protein 70 (Hsp70), mitochondrial and cytosolic manganese superoxide dismutase (mit-MnSOD and cyt-MnSOD) (Brouwer et al 2003), hemocyanin 1 (Brouwer et al 2002), Cu and Cd metallothionein (CuMT3 and CdMT1) (Syring et al 2000), beta-actin, and ribosomal proteins S15, L23 and S20, were cloned and sequenced from hepatopancreas tissue of blue crab. The blue crab genes were PCRamplified and then robotically spotted in duplicate onto neutral nylon membranes together with various controls, including exogenous Arabidopsis 'spiking' genes, as described elsewhere (Larkin et al 2003).…”

Section: Methodsmentioning

confidence: 99%

Molecular indicators of hypoxia in the blue crab Callinectes sapidus

Brown‐Peterson¹,

Larkin²,

Denslow³

et al. 2005

Mar. Ecol. Prog. Ser.

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Occurrence of hypoxia in estuarine waters is increasing, and recovery of impacted estuaries is slow. Detection of early effects of hypoxia is needed for timely remedial action to be taken. Here we examine the use of hypoxia-responsive gene and protein expression profiles in the blue crab Callinectes sapidus as early indicators of impacts of hypoxia. We cloned 23 potential hypoxiaresponsive genes, which were used to construct gene macroarrays. Crabs exposed to chronic hypoxia (2.5 ppm dissolved oxygen [DO]) for 15 d showed significant (p < 0.05) changes in gene expression of heat shock protein 70 (Hsp70), copper metallothionein (CuMt3), cytosolic MnSOD (cyt-MnSOD) and ribosomal proteins S15 and L23. In all cases except for CuMt3, gene expression decreased after 5 d exposure to hypoxia. Expression of Hsp70, CuMt3 and cyt-MnSOD also increased (p < 0.05) in normoxic crabs held for 15 d, suggesting confounding effects from confinement stress. Hemocyanin protein concentrations changed significantly (p = 0.005) across the 15 d chronic hypoxia exposure. Hemocyanin in crabs exposed to 10 d intermittent hypoxia (2.5 to 8 ppm DO over a 24 h cycle) did not change, but cyt-MnSOD gene expression increased significantly (p = 0.037), whereas cytochrome c oxidase subunit 1 (ccox1) showed a 2.2-fold downregulation. Blue crabs collected from Pensacola Bay, Florida, showed significant (p < 0.006) downregulation of ccox1 and cyt-MnSOD gene expression as well as hemocyanin protein levels at a diurnally hypoxic marsh site. Several hypoxiaresponsive genes (elongation factor 2, cryptocyanin, hemocyanin) were also significantly elevated (p < 0.006) in intermolt versus premolt normoxic crabs. The identification of hypoxia-responsive genes and proteins in the blue crab is a promising first step towards the development of sensitive molecular tools for the detection of sublethal effects of hypoxia in estuarine-resident species.KEY WORDS: Dissolved oxygen · Hypoxia · Crustacea · Molecular biomarker · Gene arrays Resale or republication not permitted without written consent of the publisher

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Section: Methodsmentioning

confidence: 99%

Molecular indicators of hypoxia in the blue crab Callinectes sapidus

Brown‐Peterson¹,

Larkin²,

Denslow³

et al. 2005

Mar. Ecol. Prog. Ser.

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“…In addition to Sod4, Sod5, and Sod6, C. albicans contains an irregular pair of cytosolic SODs that use either Cu (Cu/Zn-Sod1) or Mn (Sod3) as a catalytic cofactor (32). The vast majority of eukaryotes have only a Cu/Zn-SOD in the cytosol, and a few rare organisms (crustaceans and photosynthetic microbes) contain a cytosolic Mn-SOD, but not Cu/Zn-SOD (38,39). C. albicans and related fungi are the only organisms known to have both.…”

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confidence: 99%

Candida albicans adapts to host copper during infection by swapping metal cofactors for superoxide dismutase

Gleason

Zhang

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

100

150

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Copper is both an essential nutrient and potentially toxic metal, and during infection the host can exploit Cu in the control of pathogen growth. Here we describe a clever adaptation to Cu taken by the human fungal pathogen Candida albicans. In laboratory cultures with abundant Cu, C. albicans expresses a Cu-requiring form of superoxide dismutase (Sod1) in the cytosol; but when Cu levels decline, cells switch to an alternative Mn-requiring Sod3. This toggling between Cu- and Mn-SODs is controlled by the Cu-sensing regulator Mac1 and ensures that C. albicans maintains constant SOD activity for cytosolic antioxidant protection despite fluctuating Cu. This response to Cu is initiated during C. albicans invasion of the host where the yeast is exposed to wide variations in Cu. In a murine model of disseminated candidiasis, serum Cu was seen to progressively rise over the course of infection, but this heightened Cu response was not mirrored in host tissue. The kidney that serves as the major site of fungal infection showed an initial rise in Cu, followed by a decline in the metal. C. albicans adjusted its cytosolic SODs accordingly and expressed Cu-Sod1 at early stages of infection, followed by induction of Mn-Sod3 and increases in expression of CTR1 for Cu uptake. Together, these studies demonstrate that fungal infection triggers marked fluctuations in host Cu and C. albicans readily adapts by modulating Cu uptake and by exchanging metal cofactors for antioxidant SODs.

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“…In crustaceans, three types of SOD: cytosolic Mn-SOD, mitochondrial Mn-SOD and extracellular Cu/Zn-SOD have been cloned from blue crab C. sapidus (Donnelly et al 1989). Decapod crustaceans have one extracellular Cu/Zn-SOD, and showed a completely different sequence from their cytosolic Mn-SOD (Brouwer et al 2003;Johansson et al 1999). Both Cu/Zn-SOD and Mn-SOD have been reported to be present in shore crab C. maenas (Orbea et al 2000) and blue crab C. sapidus (Brouwer et al 2003).…”

Section: Effect Of Ph On Enzyme Activity and Stabilitymentioning

confidence: 99%

“…Decapod crustaceans have one extracellular Cu/Zn-SOD, and showed a completely different sequence from their cytosolic Mn-SOD (Brouwer et al 2003;Johansson et al 1999). Both Cu/Zn-SOD and Mn-SOD have been reported to be present in shore crab C. maenas (Orbea et al 2000) and blue crab C. sapidus (Brouwer et al 2003).…”

Section: Effect Of Ph On Enzyme Activity and Stabilitymentioning

confidence: 99%

A heat-stable Cu/Zn superoxide dismutase from the viscera of sardinelle (Sardinella aurita): purification and biochemical characterization

et al. 2014

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Superoxide dismutase (SOD) is an enzyme that protects against oxidative stress from superoxide radicals in living cells. This enzyme was extracted from sardinelle (Sardinella aurita) viscera, purified and characterized. The Cu/Zn-SOD was purified to homogeneity by the three-step procedure consisting of the heating at 65• C for 15 min, precipitation with ammonium sulphate (30-60%, w/v) and Sephadex G-100 gel filtration with a 7.17-fold increase in specific activity. The molecular weight of the native enzyme was estimated to be 40 kDa by G-125 gel filtration on HPLC column and that of the subunit mass, deduced by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, was 20 kDa. Thus the native enzyme appeared to be a homodimer. The optimum pH and temperature for the purified SOD activity were determined to be pH 7.0 and 40• C, respectively. It retained more than 85% of its initial activity after 1 h of incubation at 50• C. The enzyme had a broad stability pH range of 6.0-9.0. The N-terminal sequence of the purified enzyme was VLKAVCVLKGTGEVT. This sequence exhibited a high degree of sequence similarity with other fish Cu/Zn SODs.

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Replacement of a cytosolic copper/zinc superoxide dismutase by a novel cytosolic manganese superoxide dismutase in crustaceans that use copper (haemocyanin) for oxygen transport

Cited by 101 publications

References 43 publications

Molecular indicators of hypoxia in the blue crab Callinectes sapidus

Molecular indicators of hypoxia in the blue crab Callinectes sapidus

Candida albicans adapts to host copper during infection by swapping metal cofactors for superoxide dismutase

A heat-stable Cu/Zn superoxide dismutase from the viscera of sardinelle (Sardinella aurita): purification and biochemical characterization

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