Restricted accessResearch articleFirst published online 2018-05
The Broad-Spectrum Antimicrobial Potential of [Mn(CO) 4 (S 2 CNMe(CH 2 CO 2 H))],a Water-Soluble CO-Releasing Molecule (CORM-401): Intracellular Accumulation,Transcriptomic and Statistical Analyses,and Membrane Polarization
Carbon monoxide (CO)-releasing molecules (CORMs) are candidates for animal and antimicrobial therapeutics. We aimed to probe the antimicrobial potential of a novel manganese CORM.
Results:
[Mn(CO)4S2CNMe(CH2CO2H)], CORM-401, inhibits growth of Escherichia coli and several antibiotic-resistant clinical pathogens. CORM-401 releases CO that binds oxidases in vivo, but is an ineffective respiratory inhibitor. Extensive CORM accumulation (assayed as intracellular manganese) accompanies antimicrobial activity. CORM-401 stimulates respiration, polarizes the cytoplasmic membrane in an uncoupler-like manner, and elicits loss of intracellular potassium and zinc. Transcriptomics and mathematical modeling of transcription factor activities reveal a multifaceted response characterized by elevated expression of genes encoding potassium uptake, efflux pumps, and envelope stress responses. Regulators implicated in stress responses (CpxR), respiration (Arc, Fnr), methionine biosynthesis (MetJ), and iron homeostasis (Fur) are significantly disturbed. Although CORM-401 reduces bacterial growth in combination with cefotaxime and trimethoprim, fractional inhibition studies reveal no interaction.
Innovation:
We present the most detailed microbiological analysis yet of a CORM that is not a ruthenium carbonyl. We demonstrate CO-independent striking effects on the bacterial membrane and global transcriptomic responses.
Conclusions:
CORM-401, contrary to our expectations of a CO delivery vehicle, does not inhibit respiration. It accumulates in the cytoplasm, acts like an uncoupler in disrupting cytoplasmic ion balance, and triggers multiple effects, including osmotic stress and futile respiration.
Rebound Track:
This work was rejected during standard peer review and rescued by rebound peer review (Antioxid Redox Signal 16: 293–296, 2012) with the following serving as open reviewers: Miguel Aon, Giancarlo Biagini, James Imlay, and Nigel Robinson. Antioxid. Redox Signal. 28, 1286–1308.
Get full access to this article
View all access options for this article.
References
1.
AlvarezAF, and GeorgellisD. In vitro and in vivo analysis of the ArcB/A redox signaling pathway. Methods Enzymol, 471: 205–228, 2010.
2.
AsifHM, RolfeMD, GreenJ, LawrenceND, RattrayM, and SanguinettiG. TFInfer: a tool for probabilistic inference of transcription factor activities. Bioinformatics, 26: 2635–2636, 2010.
3.
BhattacharjeeR, MohamedMR, SahaM, SolisKP, MayerR, BarrettP, AlHasanI, AboalsamhG, CepinskasG, and LukeP. Warm perfusion of carbon monoxide releasing molecule 401 reduces inflammation and damage in preclinical donation after cardiac death kidney transplantation model. J Urol, 195: E430–E430, 2016.
4.
BogachevAV, MurtazinaRA, ShestopalovAI, and SkulachevVP. Induction of the Escherichia coli cytochrome d by low ΔμH+ and by sodium ions. Eur J Biochem, 232: 304–308, 1995.
5.
BongaertsJ, ZoskeS, WeidnerU, and UndenG. Transcriptional regulation of the proton translocating NADH dehydrogenase genes (nuoA-N) of Escherichia coli by electron acceptors, electron donors and gene regulators. Mol Microbiol, 16: 521–534, 1995.
6.
CastorLN, and ChanceB. Photochemical action spectra of carbon monoxide-inhibited respiration. J Biol Chem, 217: 453–465, 1955.
7.
CrookSH, MannBE, MeijerA, AdamsH, SawleP, ScapensD, and MotterliniR. Mn(CO)4{S2CNMe(CH2CO2H)}, a new water-soluble CO-releasing molecule. Dalton Trans, 40: 4230–4235, 2011.
8.
DavidgeKS, MotterliniR, MannBE, WilsonJL, and PooleRK. Carbon monoxide in biology and microbiology: surprising roles for the “Detroit perfume”. Adv Microb Physiol, 56: 85–167, 2009.
9.
DavidgeKS, SanguinettiG, YeeCH, CoxAG, McLeodCW, MonkCE, MannBE, MotterliniR, and PooleRK. Carbon monoxide-releasing antibacterial molecules target respiration and global transcriptional regulators. J Biol Chem, 284: 4516–4524, 2009.
10.
DegnH, LilleorM, and IversenJJL. The occurrence of a stepwise-decreasing respiration rate during oxidative assimilation of different substrates by resting Klebsiella aerogenes in a system open to oxygen. Biochem J, 136: 1097–1104, 1973.
11.
DegnH, LundsgaardJS, PetersenLC, and OrmickiA. Polarographic measurement of steady state kinetics of oxygen uptake by biochemical samples. Meth Biochem Anal, 26: 47–77, 2006.
12.
DesmardM, DavidgeKS, BouvetO, MorinD, RouxD, ForestiR, RicardJD, DenamurE, PooleRK, MontraversP, MotterliniR, and BoczkowskiJ. A carbon monoxide-releasing molecule (CORM-3) exerts bactericidal activity against Pseudomonas aeruginosa and improves survival in an animal model of bacteraemia. FASEB J, 23: 1023–1031, 2009.
13.
DeWulfP, KwonO, and LinECC. The CpxRA signal transduction system of Escherichia coli: growth-related autoactivation and control of unanticipated target operons. J Bacteriol, 181: 6772–6778, 1999.
14.
DorelC, VidalO, Prigent-CombaretC, ValletI, and LejeuneP. Involvement of the Cpx signal transduction pathway of E. coli in biofilm formation. FEMS Microbiol Lett, 178: 169–175, 1999.
15.
EjimL, FarhaMA, FalconerSB, WildenhainJ, CoombesBK, TyersM, BrownED, and WrightGD. Combinations of antibiotics and nonantibiotic drugs enhance antimicrobial efficacy. Nat Chem Biol, 7: 348–350, 2011.
16.
Fayad-KobeissiS, RatovonantenainaJ, DabireH, WilsonJL, RodriguezAM, BerdeauxA, Dubois-RandeJL, MannBE, MotterliniR, and ForestiR. Vascular and angiogenic activities of CORM-401, an oxidant-sensitive CO-releasing molecule. Biochem Pharmacol, 102: 64–77, 2016.
17.
FewsonCA, PooleRK, and ThurstonCF. Spectrophotometry in microbiology: symbols and terminology, scattered thoughts on opaque problems. Soc Gen Microbiol Q, 11: 87–89, 1984.
18.
FukutoJM, CarringtonSJ, TantilloDJ, HarrisonJG, IgnarroLJ, FreemanBA, ChenA, and WinkDA. Small molecule signaling agents: the integrated chemistry and biochemistry of nitrogen oxides, oxides of carbon, dioxygen, hydrogen sulfide, and their derived species. Chem Res Toxicol, 25: 769–793, 2012.
19.
GiamarellouH, AntoniadouA, and KanellakopoulouK. Acinetobacter baumannii: a universal threat to public health?. Int J Antimicrob Agents, 32: 106–119, 2008.
20.
GillEE, FrancoOL, and HancockRE. Antibiotic adjuvants: diverse strategies for controlling drug-resistant pathogens. Chem Biol Drug Des, 85: 56–78, 2015.
21.
GovanJR, and DereticV. Microbial pathogenesis in cystic fibrosis: mucoid Pseudomonas aeruginosa and Burkholderia cepacia. Microbiol Rev, 60: 539–574, 1996.
22.
GovanJR, and FyfeJA. Mucoid Pseudomonas aeruginosa and cystic fibrosis: resistance of the mucoid from to carbenicillin, flucloxacillin and tobramycin and the isolation of mucoid variants in vitro. J Antimicrob Chemother, 4: 233–240, 1978.
23.
GrahamAI, SanguinettiG, BramallN, McLeodCW, and PooleRK. Dynamics of a starvation-to-surfeit shift: a transcriptomic and modelling analysis of the bacterial response to zinc reveals transient behaviour of the Fur and SoxS regulators. Microbiology, 158: 284–292, 2012.
24.
GreieJC. The KdpFABC complex from Escherichia coli: a chimeric K+ transporter merging ion pumps with ion channels. Eur J Cell Biol, 90: 705–710, 2011.
25.
IuchiS, ChepuriV, FuHA, GennisRB, and LinECC. Requirement for terminal cytochromes in generation of the aerobic signal for the arc regulatory system in Escherichia coli: study utilizing deletions and lac fusions of cyo and cyd. J Bacteriol, 172: 6020–6025, 1990.
26.
IuchiS, and LinECC. Adaptation of Escherichia coli to redox environments by gene expression. Mol Microbiol, 9: 9–15, 1993.
27.
JesseHE, NyeTL, McLeanS, GreenJ, MannBE, and PooleRK. The terminal oxidase cytochrome bd-I in Escherichia coli has lower susceptibility than cytochromes bd-II or bo' to inhibition by the carbon monoxide-releasing molecule, CORM-3: N-acetylcysteine reduces CO-RM uptake and inhibition of respiration. Biochim Biophys Acta, 1834: 1693–1703, 2013.
28.
KaczaraP, MotterliniR, RosenGM, AugustynekB, BednarczykP, SzewczykA, ForestiR, and ChlopickiS. Carbon monoxide released by CORM-401 uncouples mitochondrial respiration and inhibits glycolysis in endothelial cells: a role for mitoBK(Ca) channels. Biochim Biophys Acta Bioenergetics, 1847: 1297–1309, 2015.
29.
KeilinD. The History of Cell Respiration and Cytochrome. Cambridge: Cambridge University Press, 1966, p. 416.
30.
KempfB, and BremerE. Uptake and synthesis of compatible solutes as microbial stress responses to high-osmolality environments. Arch Microbiol, 170: 319–330, 1998.
31.
KobeissiSF, WilsonJL, MichelB, Dubois-RandeJ-L, MotterliniR, and ForestiR. Pharmacological activities of CORM-401, a redox-sensitive carbon monoxide-releasing molecule, in H9C2 cardiomycetes. Archiv Cardiovasc Dis, 6Suppl 1: 17, 2014.
32.
KochAL. Growth measurement. In: Methods for General and Molecular Bacteriology, edited by GerhardtP, MurrayRGE, WoodWA, KriegNR. Washington, DC: American Society for Microbiology, 1994, pp. 248–277.
33.
LaermannV, CudicE, KipschullK, ZimmannP, and AltendorfK. The sensor kinase KdpD of Escherichia coli senses external K+. Mol Microbiol, 88: 1194–1204, 2013.
34.
LaiminsLA, RhoadsDB, and EpsteinW. Osmotic control of kdp operon expression in Escherichia coli. Proc Natl Acad Sci U S A, 78: 464–468, 1981.
35.
LefflerCW, ParfenovaH, and JaggarJH. Carbon monoxide as an endogenous vascular modulator. Am J Physiol Heart Circ Physiol, 301: H1–H11, 2011.
36.
Lo IaconoL, BoczkowskiJ, ZiniR, SalouageI, BerdauxA, MotterliniR, and MorinD. A carbon monoxide-releasing molecule (CORM-3) uncouples mitochondrial respiration and modulates the production of reactive oxygen species. Free Rad Biol Med, 50: 1556–1564, 2011.
37.
MannBE. CO-Releasing Molecules: a Personal View. Organometallics, 31: 5728–5735, 2012.
38.
McLeanS, BeggR, JesseHE, MannBE, SanguinettiG, and PooleRK. Analysis of the bacterial response to Ru(CO)3Cl(glycinate) (CORM-3) and the inactivated compound identifies the role played by the ruthenium compound and reveals sulfur-containing species as a major target of CORM-3 action. Antioxid Redox Signal, 19: 1999–2012, 2013.
39.
McLeanS, MannBE, and PooleRK. Sulfite species enhance carbon monoxide release from CO-releasing molecules: implications for the deoxymyoglobin assay of activity. Anal Biochem, 427: 36–40, 2012.
NagelC, McLeanS, PooleRK, BraunschweigH, KramerT, and SchatzschneiderU. Introducing [Mn(CO)3(tpa-k3N)]+ as a novel photoactivatable CO-releasing molecule with well-defined iCORM intermediates—synthesis, spectroscopy, and antibacterial activity. Dalton Trans, 43: 9986–9997, 2014.
42.
NaseemR, HollandIB, JacqA, WannKT, and CampbellAK. pH and monovalent cations regulate cytosolic free Ca2+ in E. coli. Biochim Biophys Acta Biomembranes, 1778: 1415–1422, 2008.
43.
NichollsDG, and FergusonSJ. Bioenergetics 3. London: Academic Press, 2002.
44.
NightingaleCH, AmbrosePG, DrusanoGL, and MurakawaT (Eds). Antimicrobial Pharmacodynamics in Theory and Clinical Practice, Second Edition (Infectious Disease and Therapy). New York and London: Informa Healthcare, 2007.
NobreLS, Al-ShahrourF, DopazoJ, and SaraivaLM. Exploring the antimicrobial action of a carbon monoxide-releasing compound through whole-genome transcription profiling of Escherichia coli. Microbiology, 155: 813–824, 2009.
47.
NomuraT, CoxCD, BaviN, SokabeM, and MartinacB. Unidirectional incorporation of a bacterial mechanosensitive channel into liposomal membranes. FASEB J, 29: 4334–4345, 2015.
48.
OddsFC. Synergy, antagonism, and what the chequerboard puts between them. J Antimicrob Chemother, 52: 1, 2003.
49.
OuttenCE, and O'HalloranTV. Femtomolar sensitivity of metalloregulatory proteins controlling zinc homeostasis. Science, 292: 2488–2492, 2001.
50.
PapapetropoulosA, ForestiR, and FerdinandyP. Pharmacology of the ‘gasotransmitters’ NO, CO and H2S: translational opportunities. Br J Pharmacol, 172: 1395–1396, 2015.
51.
PatzerSI, and HantkeK. The ZnuABC high-affinity zinc uptake system and its regulator Zur in Escherichia coli. Mol Microbiol, 28: 1199–1210, 1998.
52.
PooleRK. The influence of growth substrate and capacity for oxidative phosphorylation on respiratory oscillations in synchronous cultures of Escherichia coli K12. J Gen Microbiol, 99: 369–377, 1977.
53.
QuanS, HinikerA, ColletJF, and BardwellJC. Isolation of bacteria envelope proteins. Methods Mol Biol, 966: 359–366, 2013.
54.
RadchenkoMV, TanakaK, WaditeeR, OshimiS, MatsuzakiY, FukuharaM, KobayashiH, TakabeT, and NakamuraT. Potassium/proton antiport system of Escherichia coli. J Biol Chem, 281: 19822–19829, 2006.
RanaN, McLeanS, MannBE, and PooleRK. Interaction of the carbon monoxide-releasing molecule Ru(CO)3Cl(glycinate) (CORM-3) with Salmonella enterica serovar Typhimurium: in situ measurements of carbon monoxide binding by integrating cavity dual-beam spectrophotometry. Microbiology, 160: 2771–2779, 2014.
57.
SanguinettiG, LawrenceND, and RattrayM. Probabilistic inference of transcription factor concentrations and gene-specific regulatory activities. Bioinformatics, 22: 2775–2781, 2006.
58.
SanguinettiG, RattrayM, and LawrenceND. A probabilistic dynamical model for quantitative inference of the regulatory mechanism of transcription. Bioinformatics, 22: 1753–1759, 2006.
59.
Santos-SilvaT, MukhopadhyayA, SeixasJD, BernardesGJL, RomaoCC, and RomaoMJ. CORM-3 reactivity toward proteins: the crystal structure of a Ru(II) dicarbonyl-lysozyme complex. J Am Chem Soc, 133: 1192–1195, 2011.
60.
SawleP, ForestiR, MannBE, JohnsonTR, GreenCJ, and MotterliniR. Carbon monoxide-releasing molecules (CO-RMs) attenuate the inflammatory response elicited by lipopolysaccharide in RAW264.7 murine macrophages. Br J Pharmacol, 145: 800–810, 2005.
61.
SimsPJ, WaggonerAS, WangCH, and HoffmanJF. Studies on the mechanism by which cyanine dyes measure membrane potential in red blood cells and phosphatidylcholine vesicles. Biochemistry, 13: 3315–3330, 1974.
62.
SoupeneE, KingN, LeeH, and KustuS. Aquaporin Z of Escherichia coli: reassessment of its regulation and physiological role. J Bacteriol, 184: 4304–4307, 2002.
63.
SouthamHM, ButlerJA, ChapmanJA, and PooleRK. The microbiology of ruthenium complexes. Adv Microb Physiol, 71:1–96, 2017.
64.
StratevaT, and YordanovD. Pseudomonas aeruginosa - a phenomenon of bacterial resistance. J Med Microbiol, 58: 1133–1148, 2009.
65.
StropP, BassR, and ReesDC. Prokaryotic mechanosensitive channels. Adv Protein Chem, 63: 177–209, 2003.
66.
SugiuraA, HirokawaK, NakashimaK, and MizunoT. Signal-sensing mechanisms of the putative osmosensor KdpD in Escherichia coli. Mol Microbiol, 14: 929–938, 1994.
67.
Tinajero-TrejoM, RanaN, NagelC, JesseHE, SmithTW, WarehamLK, HipplerM, SchatzschneiderU, and PooleRK. Antimicrobial activity of the manganese photoactivated carbon monoxide-releasing molecule Mn(CO)3(tpa-kappa n-3)+ against a pathogenic Escherichia coli that causes urinary infections. Antioxid Redox Signal, 24: 765–780, 2016.
68.
TotsikaM, BeatsonSA, SarkarS, PhanMD, PettyNK, BachmannN, SzubertM, SidjabatHE, PatersonDL, UptonM, and SchembriMA. Insights into a multidrug resistant Escherichia coli pathogen of the globally disseminated ST131 lineage: genome analysis and virulence mechanisms. PLoS One, 6: e26578, 2011.
69.
WarehamLK, BeggR, JesseHE, van BeilenJWA, AliS, SvistunenkoD, McLeanS, HellingwerfKJ, SanguinettiG, and PooleRK. Carbon monoxide gas is not inert, but global, in its consequences for bacterial gene expression, iron acquisition, and antibiotic resistance. Antioxid Redox Signal, 24: 1013–1028, 2016.
70.
WarehamLK, PooleRK, and Tinajero-TrejoM. CO-releasing metal carbonyl compounds as antimicrobial agents in the post-antibiotic era. J Biol Chem, 290: 18999–19007, 2015.
71.
WHO. 2017. Global priority list of antibiotic-resistant bacteria to guide research, discovery, and development of new antibiotics. www.who.int/medicines/publications/global-priority-list-antibiotic-resistant-bacteria/en (accessed February27, 2017).
72.
WilsonJL, BouillaudF, AlmeidaAS, VieiraHL, OuidjaMO, Dubois-RandeJL, ForestiR, and MotterliniR. Carbon monoxide reverses the metabolic adaptation of microglia cells to an inflammatory stimulus. Free Rad Biol Med, 104: 311–323, 2017.
73.
WilsonJL, JesseHE, HughesBM, LundV, NaylorK, DavidgeKS, CookGM, MannBE, and PooleRK. Ru(CO)3Cl(glycinate) (CORM-3): a CO-releasing molecule with broad-spectrum antimicrobial and photosensitive activities against respiration and cation transport in Escherichia coli. Antioxid Redox Signal, 19: 497–509, 2013.
74.
WilsonJL, WarehamLK, McLeanS, BeggR, GreavesS, MannBE, SanguinettiG, and PooleRK. CO-Releasing molecules have nonheme targets in bacteria: transcriptomic, mathematical modeling and biochemical analyses of CORM-3 [Ru(CO)Cl(glycinate)] Actions on a heme-deficient mutant of Escherichia coli. Antioxid Redox Signal, 23: 148–162, 2015.
75.
WoodJM. Osmosensing by bacteria: signals and membrane-based sensors. Microbiol Mol Biol Rev, 63: 230–262, 1999.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
