Abstract
Metal ions can be both essential components of cells as well as potential toxins if present in excess. Organisms utilize a variety of protein systems to maintain the concentration of metal ions within the appropriate range for cellular function, and to avoid concentrations where cellular damage can occur. In bacteria, numerous proteins contribute to copper homeostasis, including copper transporters, chelators, and redox enzymes. The genes that encode these proteins are often found in clusters, thus providing modular components that work together to achieve homeostasis. A better understanding of how these components function and cooperate to achieve metal ion resistance is needed, given the extensive use of metal ions, including copper, as broad-spectrum biocides in a variety of clinical and environmental settings. The copG gene is a common component of such copper resistance clusters, but its contribution to copper resistance is not well understood. In this review the available information about the CopG protein encoded by this gene is summarized. Comparison of the recent structure to diverse copper-containing metallochaperones, metalloenzymes, and electron transfer proteins suggests that CopG is a redox enzyme that uses multiple copper ions as active site redox cofactors to act on additional copper ion substrates. Mechanisms for both oxidase and reductase activity are proposed, and the biological advantages that these activities can contribute in conjunction with existing systems are described.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Arciero DM, Pierce BS, Hendrich MP, Hooper AB (2002) Nitrosocyanin, a Red Cupredoxin-like Protein from Nitrosomonas europaea. Biochemistry 41(6). https://doi.org/10.1021/bi015908w
Arguello JM (2003) Identification of ion-selectivity determinants in heavy-metal transport P-1B-type ATPases. J Membr Biol 195(2). https://doi.org/10.1007/s00232-003-2048-2
Asiani KR, Williams H, Bird L, Jenner M, Searle MS, Hobman JL, Scott DJ, Soultanas P (2016) SilE is an intrinsically disordered periplasmic “molecular sponge” involved in bacterial silver resistance. Molecular Microbiology 101(5). https://doi.org/10.1111/mmi.13399
Atkinson HJ, Babbitt PC (2009) An Atlas of the Thioredoxin Fold Class Reveals the Complexity of Function-Enabling Adaptations. PLoS Comput Biol 5(10):e1000541. https://doi.org/10.1371/journal.pcbi.1000541. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2757866/
Augustine AJ, Kjaergaard C, Qayyum M, Ziegler L, Kosman DJ, Hodgson KO, Hedman B, Solomon EI (2010) Systematic perturbation of the trinuclear copper cluster in the multicopper oxidases: The role of active site asymmetry in its reduction of O2 to H2O. J Am Chem Soc 132(17). https://doi.org/10.1021/ja909143d
Bagai I, Rensing C, Blackburn NJ, McEvoy MM (2008) Direct metal transfer between periplasmic proteins identifies a bacterial copper chaperone. Biochemistry 47(44). https://doi.org/10.1021/bi801638m
Banaszak K, Mechin I, Frost G, Rypniewski W (2004) Structure of the reduced disulfide-bond isomerase DsbC from Escherichia coli. Acta Crystallogr D 60(10). https://doi.org/10.1107/S0907444904018359. https://scripts.iucr.org/cgi-bin/paper?hm5018
Barkay T, Miller SM, Summers AO (2003) Bacterial mercury resistance from atoms to ecosystems. Fems Microbiol Rev 27(2-3). https://doi.org/10.1016/S0168-6445(03)00046-9
Beinert H (2000) Iron-sulfur proteins: ancient structures, still full of surprises. JBIC J Biol Inorg Chem 5(1):2–15. https://doi.org/10.1007/s007750050002. http://link.springer.com/10.1007/s007750050002
Beinert H, Holm RH, Munck E (1997) Iron-sulfur clusters: Nature’s modular, multipurpose structures. Science 277(5326):653–659. https://doi.org/10.1126/science.277.5326.653. https://www.science.org/doi/10.1126/science.277.5326.653
Beswick P, Hall G, Hook A, Little K, Mcbrien D, Lott K (1976) Copper toxicity - evidence for conversion of cupric to cuprous copper invivo under anaerobic conditions. Chem Biol Interact 14(3-4). https://doi.org/10.1016/0009-2797(76)90113-7
Bewley KD, Dey M, Bjork RE, Mitra S, Chobot SE, Drennan CL, Elliott SJ (2015) Rheostat re-wired: Alternative hypotheses for the control of thioredoxin reduction potentials. PLOS ONE 10(4). https://doi.org/10.1371/journal.pone.0122466. https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0122466
Blackburn NJ, Barr ME, Woodruff WH, van der Ooost J, de Vries S (1994) Metal-metal bonding in biology: EXAFS Evidence for a 2.5 .ANG. Copper-copper bond in the CuA center of cytochrome oxidase. Biochemistry 33(34). https://doi.org/10.1021/bi00200a022
Bruins MR, Kapil S, Oehme FW (2000) Microbial resistance to metals in the environment. Ecotoxicol Environ Saf 45(3). https://doi.org/10.1006/eesa.1999.1860
Chacon KN, Mealman TD, McEvoy MM, Blackburn NJ (2014) Tracking metal ions through a Cu/Ag efflux pump assigns the functional roles of the periplasmic proteins. Proc Natl Acad Sci 111(43). https://doi.org/10.1073/pnas.1411475111. https://www.pnas.org/doi/full/10.1073/pnas.1411475111
Chivers PT, Prehoda KE, Raines RT (1997) The CXXC Motif: A rheostat in the active site. Biochemistry 36(14):4061–4066. https://doi.org/10.1021/bi9628580. https://pubs.acs.org/doi/10.1021/bi9628580
Denoncin K, Collet JF (2013) Disulfide bond formation in the bacterial periplasm: major achievements and challenges ahead. Antioxid Redox Signaling 19(1):63–71. https://doi.org/10.1089/ars.2012.4864. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3676657/
Durand A, Fouesnard M, Bourbon ML, Steunou AS, Lojou E, Dorlet P, Ouchane S (2021) A periplasmic cupredoxin with a green CuT1.5 center is involved in bacterial copper tolerance. Metallomics 13(12):mfab067. https://doi.org/10.1093/mtomcs/mfab067. https://academic.oup.com/metallomics/article/doi/10.1093/mtomcs/mfab067/6428408
El-Gebali S, Mistry J, Bateman A, Eddy SR, Luciani A, Potter SC, Qureshi M, Richardson LJ, Salazar GA, Smart A, Sonnhammer ELL, Hirsh L, Paladin L, Piovesan D, Tosatto SCE, Finn RD (2019) The Pfam protein families database in 2019. Nucl Acids Res 47(D1). https://doi.org/10.1093/nar/gky995
Giachino A, Waldron KJ (2020) Copper tolerance in bacteria requires the activation of multiple accessory pathways. Molecular Microbiology 114(3). https://doi.org/10.1111/mmi.14522
Grass G, Rensing C (2001) CueO is a multi-copper oxidase that confers copper tolerance in Escherichia coli. Biochem Biophys Res Commun 286(5). https://doi.org/10.1006/bbrc.2001.5474
Guddat LW, Bardwell JC, Glockshuber R, Huber-Wunderlich M, Zander T, Martin JL (1997) Structural analysis of three His32 mutants of DsbA: support for an electrostatic role of His32 in DsbA stability. Protein Science 6(9):1893–1900. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2143798/
Guddat LW, Bardwell JC, Martin JL (1998) Crystal structures of reduced and oxidized DsbA: investigation of domain motion and thiolate stabilization. Structure 6(6):757–767. https://doi.org/10.1016/S0969-2126(98)00077-X. https://www.sciencedirect.com/science/article/pii/S096921269800077X
Gupta A, Matsui K, Lo JF, Silver S (1999) Molecular basis for resistance to silver cations in Salmonella. Nature Medicine 5(2). https://doi.org/10.1038/5545
Haebel PW, Goldstone D, Katzen F, Beckwith J, Metcalf P (2002) The disulfide bond isomerase DsbC is activated by an immunoglobulin-fold thiol oxidoreductase: crystal structure of the DsbC/DsbDα complex. EMBO J 21(18):4774–4784. https://doi.org/10.1093/emboj/cdf489. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC126285/
Halsted TP, Yamashita K, Gopalasingam CC, Shenoy RT, Hirata K, Ago H, Ueno G, Blakeley MP, Eady RR, Antonyuk SV, Yamamoto M, Hasnain SS (2019) Catalytically important damage-free structures of a copper nitrite reductase obtained by femtosecond X-ray laser and room-temperature neutron crystallography. IUCrJ 6. https://doi.org/10.1107/S2052252519008285
Hausrath AC, Ramirez NA, Ly AT, McEvoy MM (2020) The bacterial copper resistance protein CopG contains a cysteine-bridged tetranuclear copper cluster. J Biol Chem 295(32):11364–11376. https://doi.org/10.1074/jbc.RA120.013907. https://linkinghub.elsevier.com/retrieve/pii/S0021925817492258
Hiniker A, Collet JF, Bardwell JCA (2005) Copper stress causes an in vivo requirement for the Escherichia coli disulfide isomerase DsbC. J Biol Chem 280(40). https://doi.org/10.1074/jbc.M505742200. https://www.jbc.org/article/S0021-9258(20)78961-1/abstract
Hol W (1985) The role of the alpha-helix dipole in protein function and structure. Prog Biophys Mol Biol 45(3). https://doi.org/10.1016/0079-6107(85)90001-X
Holmgren A (1985) Thioredoxin. Annu Rev Biochem 54:237–271. https://doi.org/10.1146/annurev.bi.54.070185.001321
Huber-Wunderlich M, Glockshuber R (1998) A single dipeptide sequence modulates the redox properties of a whole enzyme family. Folding Des 3(3):161–171. https://doi.org/10.1016/S1359-0278(98)00024-8, https://www.sciencedirect.com/science/article/pii/S1359027898000248
Katti S, Lemaster D, Eklund H (1990) Crystal-structure of thioredoxin from Escherichia-coli at 1.68 angstrom resolution. J Mol Biol 212(1). https://doi.org/10.1016/0022-2836(90)90313-B
Kim EH, Rensing C, McEvoy MM (2010) Chaperone-mediated copper handling in the periplasm. Nat Prod Rep 27(5). https://doi.org/10.1039/b906681k
King JD, McIntosh CL, Halsey CM, Lada BM, Niedzwiedzki DM, Cooley JW, Blankenship RE (2013) Metalloproteins diversified: The auracyanins are a family of cupredoxins that stretch the spectral and redox limits of blue copper proteins. Biochemistry 52(46). https://doi.org/10.1021/bi401163g
LaCroix LB, Shadle SE, Wang Y, Averill BA, Hedman B, Hodgson KO, Solomon EI (1996) Electronic structure of the perturbed blue copper site in nitrite reductase: spectroscopic properties, bonding, and implications for the entatic/rack state. J Am Chem Soc 118(33). https://doi.org/10.1021/ja961217p
Lieberman RL, Arciero DM, Hooper AB, Rosenzweig AC (2001) Crystal structure of a novel red copper protein from nitrosomonas europaea. Biochemistry 40(19). https://doi.org/10.1021/bi0102611
Liu J, Chakraborty S, Hosseinzadeh P, Yu Y, Tian S, Petrik I, Bhagi A, Lu Y (2014) Metalloproteins containing cytochrome, iron-sulfur, or copper redox centers. Chemical Reviews 114(8):4366–4469. https://doi.org/10.1021/cr400479b. https://pubs.acs.org/doi/10.1021/cr400479b
Macomber L, Imlay JA (2009) The iron-sulfur clusters of dehydratases are primary intracellular targets of copper toxicity. Proc Natl Acad Sci USA 106(20):8344–8349. https://doi.org/10.1073/pnas.0812808106
Malojcic G, Owen RL, Grimshaw JP, Glockshuber R (2008) Preparation and structure of the charge-transfer intermediate of the transmembrane redox catalyst DsbB. FEBS Letters 582(23-24). https://doi.org/10.1016/j.febslet.2008.07.063. https://onlinelibrary.wiley.com/doi/abs/10.1016/j.febslet.2008.07.063
Marrero K, Sanchez A, Gonzalez LJ, Ledon T, Rodriguez-Ulloa A, Castellanos-Serra L, Perez C, Fando R (2012) Periplasmic proteins encoded by VCA0261-0260 and VC2216 genes together with copA and cueR products are required for copper tolerance but not for virulence in Vibrio cholerae. Microbiology-Sgm 158. https://doi.org/10.1099/mic.0.059345-0
McCarthy AA, Haebel PW, Torronen A, Rybin V, Baker EN, Metcalf P (2000) Crystal structure of the protein disulfide bond isomerase, DsbC, from Escherichia coli. Nat Struct Biol 7(3). https://doi.org/10.1038/73295. https://www.nature.com/articles/nsb0300_196
Mealman TD, Zhou M, Affandi T, Chacon KN, Aranguren ME, Blackburn NJ, Wysocki VH, McEvoy MM (2012) N-terminal region of CusB is sufficient for metal binding and metal transfer with the metallochaperone CusF. Biochemistry 51(34). https://doi.org/10.1021/bi300596a
Nar H, Messerschmidt A, Huber R, Vandekamp M, Canters G (1991) Crystal-structure analysis of oxidized pseudomonas-aeruginosa azurin at pH 5.5 and pH 9.0 - a pH-induced conformational transition involves a peptide-bond flip. J Mol Biol 221(3). https://doi.org/10.1016/0022-2836(91)80173-R
Outten FW, Huffman DL, Hale JA, O’Halloran TV (2001) The independent cue and cus systems confer copper tolerance during aerobic and anaerobic growth in Escherichia coli. J Biol Chem 276(33). https://doi.org/10.1074/jbc.M104122200
Pal C, Asiani K, Arya S, Rensing C, Stekel DJ, Larsson DGJ, Hobman JL (2017) Metal resistance and its association with antibiotic resistance. In: Poole RK (ed) Microbiology of metal ions, vol 70. Academic Press Ltd-Elsevier Science Ltd, London. https://doi.org/10.1016/bs.ampbs.2017.02.001. https://www.webofscience.com/wos/woscc/full-record/WOS:000414265600008
Pearson R (1963) Hard and soft acids and bases. J Am Chem Soc 85(22). https://doi.org/10.1021/ja00905a001
Rensing C, Grass G (2003) Escherichia coli mechanisms of copper homeostasis in a changing environment. Fems Microbiol Rev 27(2-3). https://doi.org/10.1016/S0168-6445(03)00049-4
Roger M, Biaso F, Castelle CJ, Bauzan M, Chaspoul F, Lojou E, Sciara G, Caffarri S, Giudici-Orticoni MT, Ilbert M (2014) Spectroscopic characterization of a green copper site in a single-domain cupredoxin. PLOS ONE 9(6). https://doi.org/10.1371/journal.pone.0098941. https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0098941
Rohr AK, Hammerstad M, Andersson KK (2013) Tuning of thioredoxin redox properties by intramolecular hydrogen bonds. PLoS ONE 8(7):e69411. https://doi.org/10.1371/journal.pone.0069411. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3720550/
Rosenzweig AC, Arguello JM (2012) Toward a molecular understanding of metal transport by P-1B-type ATPases. Metal Transporters 69. https://doi.org/10.1016/B978-0-12-394390-3.00005-7
Rozhkova A, Stirnimann CU, Frei P, Grauschopf U, Brunisholz R, Grutter MG, Capitani G, Glockshuber R (2004) Structural basis and kinetics of inter- and intramolecular disulfide exchange in the redox catalyst DsbD. Embo J 23(8). https://doi.org/10.1038/sj.emboj.7600178
Ruprecht J, Iwata S, Rothery RA, Weiner JH, Maklashina E, Cecchini G (2011) Perturbation of the quinone-binding site of complex II alters the electronic properties of the proximal [3Fe-4S] iron-sulfur cluster. J Biol Chem 286(14). https://doi.org/10.1074/jbc.M110.209874
Singh SK, Grass G, Rensing C, Montfort WR (2004) Cuprous oxidase activity of CueO from Escherichia coli. J Bacteriol 186(22). https://doi.org/10.1128/JB.186.22.7815-7817.2004
Singh SK, Roberts SA, McDevitt SF, Weichsel A, Wildner GF, Grass GB, Rensing C, Montfort WR (2011) Crystal structures of multicopper oxidase CueO bound to Copper(I) and Silver(I): Functional role of a methionine-rich sequence. J Biol Chem 286(43). https://doi.org/10.1074/jbc.M111.293589
Solomon EI (2006) Spectroscopic methods in bioinorganic chemistry: Blue to green to red copper sites. Inorganic Chemistry 45(20). https://doi.org/10.1021/ic060450d
Solomon EI, Szilagyi RK, DeBeer George S, Basumallick L (2004) Electronic structures of metal sites in proteins and models: contributions to function in blue copper proteins. Chemical Reviews 104(2). https://doi.org/10.1021/cr0206317
Solomon EI, Xie X, Dey A (2008) Mixed valent sites in biological electron transfer. Chem Soc Rev 37(4). https://doi.org/10.1039/b714577m
Staehlin BM, Gibbons JG, Rokas A, O’Halloran TV, Slot JC (2016) Evolution of a heavy metal homeostasis/resistance island reflects increasing copper stress in enterobacteria. Genome Biol Evol 8(3). https://doi.org/10.1093/gbe/evw031
Suzuki S, Kataoka K, Yamaguchi K, Inoue T, Kai Y (1999) Structure/function relationships of copper-containing nitrite reductases. Coord Chem Rev 190-192:245–265. https://doi.org/10.1016/S0010-8545(99)00069-7. https://linkinghub.elsevier.com/retrieve/pii/S0010854599000697
von Rozycki T, Nies DH (2009) Cupriavidus metallidurans: evolution of a metal-resistant bacterium. Antonie Van Leeuwenhoek Int J Gen Mol Microbiol 96(2). https://doi.org/10.1007/s10482-008-9284-5
Wouters MA, Fan SW, Haworth NL (2010) Disulfides as redox switches: From molecular mechanisms to functional significance. Antioxid Redox Signal 12(1):53–91. https://doi.org/10.1089/ars.2009.2510. http://www.liebertpub.com/doi/10.1089/ars.2009.2510
Wright BW, Kamath KS, Krisp C, Molloy MP (2019) Proteome profiling of Pseudomonas aeruginosa PAO1 identifies novel responders to copper stress. BMC Microbiology 19. https://doi.org/10.1186/s12866-019-1441-7
Yano N, Muramoto K, Shimada A, Takemura S, Baba J, Fujisawa H, Mochizuki M, Shinzawa-Itoh K, Yamashita E, Tsukihara T, Yoshikawa S (2016) The Mg2+-containing water cluster of mammalian cytochrome c oxidase collects four pumping proton equivalents in each catalytic cycle. J Biol Chem 291(46). https://doi.org/10.1074/jbc.M115.711770
Zhang L, Koay M, Maher MJ, Xiao Z, Wedd AG (2006) Intermolecular transfer of copper ions from the CopC protein of Pseudomonas syringae. Crystal structures of fully loaded Cu(I)Cu(II) forms. J Am Chem Soc 128(17):5834–5850. https://doi.org/10.1021/ja058528x
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2023 Springer Nature Switzerland AG
About this chapter
Cite this chapter
Hausrath, A.C., McEvoy, M.M. (2023). Structural Analyses of the Multicopper Site of CopG Support a Role as a Redox Enzyme. In: Atassi, M.Z. (eds) Protein Reviews. Advances in Experimental Medicine and Biology(), vol 1414. Springer, Cham. https://doi.org/10.1007/5584_2022_753
Download citation
DOI: https://doi.org/10.1007/5584_2022_753
Published:
Publisher Name: Springer, Cham
Print ISBN: 978-3-031-28669-8
Online ISBN: 978-3-031-28670-4
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)