{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T13:03:40Z","timestamp":1788959020469,"version":"build-2803163510"},"reference-count":39,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2025,4,10]],"date-time":"2025-04-10T00:00:00Z","timestamp":1744243200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The discovery of carbon monoxide releasing molecules (CORMs) was one of the most impactful innovations in biochemistry, affecting multiple disciplines for the past few decades. Sixteen years ago, a ruthenium dimer-containing CORM, CORM-2, enhanced coagulation and diminished fibrinolysis in human plasma by modulation of fibrinogen, plasmin, and \u03b12-antiplasmin via CO binding to putative heme groups attached to these proteins. This finding linked CO exposure in settings involving heme oxygenase-1 upregulation during inflammation or environmental exposure to thromboembolic disease in hundreds of subsequent manuscripts. However, CO-independent effects of CORM-2 involving a putative ruthenium radical (Ru\u2022) formed during CO release was found to be responsible for many of effects by CORM-2 in other works. Using a novel approach with human plasmatic coagulation kinetic methods, Ru\u2022 was posited to bind to critical histidines and other amino acids to modulate function, and excess histidine to quench CORM-2-mediated effects. This paradigm of histidine addition would definitively address if CO or Ru\u2022 was responsible for CORM-2-mediated effects. Thus, plasma coagulation\/fibrinolytic kinetic data were assessed via thrombelastography \u00b1CORM-2, \u00b1histidine added. Histidine nearly completely abrogated CORM-2-mediated hypercoagulation in a concentration-dependent fashion; further, histidine also nearly eliminated all kinetic effects on fibrinolysis. In conclusion, CORM-2 Ru\u2022 formation, not CO release, is the true molecular mechanism modulating coagulation and fibrinolysis.<\/jats:p>","DOI":"10.3390\/ijms26083567","type":"journal-article","created":{"date-parts":[[2025,4,10]],"date-time":"2025-04-10T07:41:51Z","timestamp":1744270911000},"page":"3567","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Carbon Monoxide or Ruthenium: Will the Real Modulator of Coagulation and Fibrinolysis Please Stand Up!"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8072-9883","authenticated-orcid":false,"given":"Vance G.","family":"Nielsen","sequence":"first","affiliation":[{"name":"Department of Anesthesiology, The University of Arizona College of Medicine, Tucson, AZ 85724, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anthony R.","family":"Abeyta","sequence":"additional","affiliation":[{"name":"Ross University School of Medicine, Bridgetown BB11037, Barbados"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"4060","DOI":"10.1021\/bi00411a023","article-title":"Ruthenium-iron hybrid hemoglobins as a model for partially liganded hemoglobin: NMR studies of their tertiary and quaternary structures","volume":"27","author":"Ishimori","year":"1988","journal-title":"Biochemistry"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"9612","DOI":"10.1039\/D4DT00087K","article-title":"Metal-based carbon monoxide releasing molecules with promising cytotoxic properties","volume":"53","author":"Mansour","year":"2024","journal-title":"Dalton Trans."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"20210181","DOI":"10.1002\/EXP.20210181","article-title":"Therapeutic gas-releasing nanomedicines with controlled release: Advances and perspectives","volume":"2","author":"Zhang","year":"2022","journal-title":"Exploration"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1007\/s41061-022-00413-6","article-title":"Water-Soluble Carbon Monoxide-Releasing Molecules (CORMs)","volume":"381","author":"Khan","year":"2022","journal-title":"Top. Curr. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"E17","DOI":"10.1161\/hh0202.104530","article-title":"Carbon monoxide-releasing molecules: Characterization of biochemical and vascular activities","volume":"90","author":"Motterlini","year":"2002","journal-title":"Circ. Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e2","DOI":"10.1161\/01.RES.0000084381.86567.08","article-title":"Cardioprotective actions by a water-soluble carbon monoxide-releasing molecule","volume":"93","author":"Clark","year":"2003","journal-title":"Circ. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1097\/MBC.0b013e32832ca3a3","article-title":"Carbon monoxide releasing molecule-2 increases the velocity of thrombus growth and strength in human plasma","volume":"20","author":"Nielsen","year":"2009","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"448","DOI":"10.1097\/MBC.0b013e32832f4335","article-title":"Carbon monoxide-releasing molecule-2 decreases fibrinolysis in human plasma","volume":"20","author":"Nielsen","year":"2009","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1097\/MBC.0b013e3283333c5d","article-title":"Carbon monoxide releasing molecule-2 decreases thick diameter fibrin fibre formation in normal and Factor XIII deficient plasmas","volume":"21","author":"Nielsen","year":"2010","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1016\/j.thromres.2013.12.004","article-title":"Carbon monoxide: Anticoagulant or procoagulant?","volume":"133","author":"Nielsen","year":"2014","journal-title":"Thromb. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"756","DOI":"10.1097\/MBC.0b013e32834c7412","article-title":"Carbon monoxide-releasing molecule-2 enhances coagulation in rabbit plasma and decreases bleeding time in clopidogrel\/aspirin-treated rabbits","volume":"22","author":"Nielsen","year":"2011","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1097\/MBC.0b013e32834ea012","article-title":"Carbon monoxide-releasing molecule-2 decreases fibrinolysis in vitro and in vivo in the rabbit","volume":"23","author":"Nielsen","year":"2012","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1097\/MBC.0b013e328345c069","article-title":"Fibrinogen is a heme-associated, carbon monoxide sensing molecule: A preliminary report","volume":"22","author":"Nielsen","year":"2011","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"712","DOI":"10.1097\/MBC.0b013e32834c73f9","article-title":"Carbon monoxide and nitric oxide modulate \u03b12-antiplasmin and plasmin activity: Role of heme","volume":"22","author":"Arkebauer","year":"2011","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1097\/MBC.0b013e328344c657","article-title":"Carbon monoxide releasing molecule-2 enhances \u03b12-antiplasmin activity","volume":"22","author":"Malayaman","year":"2011","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1213\/00000539-200102000-00006","article-title":"Nitric oxide decreases coagulation protein function in rabbits as assessed by thromboelastography","volume":"92","author":"Nielsen","year":"2001","journal-title":"Anesth. Analg."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1097\/MBC.0000000000000602","article-title":"Exposure of fibrinogen and thrombin to nitric oxide donor ProliNONOate affects fibrin clot properties","volume":"28","author":"Helms","year":"2017","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"801","DOI":"10.1097\/MBC.0000000000000146","article-title":"Comparison of the effects of CORM-2, CORM-3 and CORM-A1 on coagulation in human plasma","volume":"25","author":"Nielsen","year":"2014","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.ijbiomac.2010.05.013","article-title":"Boron stabilizes peroxide mediated changes in the structure of heme proteins","volume":"47","author":"Ali","year":"2010","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.ejphar.2017.10.006","article-title":"CO-independent modification of K+ channels by tricarbonyldichlororuthenium(II) dimer (CORM-2)","volume":"815","author":"Gessner","year":"2017","journal-title":"Eur. J. Pharmacol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/0014-5793(75)80242-0","article-title":"Complete amino acid sequence of human serum albumin","volume":"58","author":"Meloun","year":"1975","journal-title":"FEBS Lett."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1007\/s11239-019-01980-0","article-title":"The anticoagulant effect of Apis mellifera phospholipase A2 is inhibited by CORM-2 via a carbon monoxide-independent mechanism","volume":"49","author":"Nielsen","year":"2020","journal-title":"J. Thromb. Thrombolysis"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Nielsen, V.G. (2024). Phosphate-Buffered Saline and Dimethyl Sulfoxide Enhance the Antivenom Action of Ruthenium Chloride against Crotalus atrox Venom in Human Plasma\u2014A Preliminary Report. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25126426"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1007\/s00775-018-1597-x","article-title":"Understanding the interactions of diruthenium anticancer agents with amino acids","volume":"23","author":"Nazarov","year":"2018","journal-title":"J. Biol. Inorg. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1111\/j.1749-6632.1972.tb16323.x","article-title":"The molecular structure of fibrinogen","volume":"202","year":"1972","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_26","first-page":"164","article-title":"Structure and function of fibrinogen","volume":"3","author":"Doolittle","year":"1977","journal-title":"Horiz. Biochem. Biophys."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"6757","DOI":"10.1016\/S0021-9258(19)44651-6","article-title":"The primary structure of human plasminogen. I. The NH 2 -terminal sequences of human plasminogen and the S-carboxymethyl heavy (A) and light (B) chain derivatives of plasmin","volume":"247","author":"Robbins","year":"1972","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1631","DOI":"10.1016\/S0021-9258(19)44237-3","article-title":"The primary structure of human plasminogen. II. The histidine loop of human plasmin: Light (B) chain active center histidine sequence","volume":"248","author":"Robbins","year":"1973","journal-title":"J. Biol. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1111\/j.1432-1033.1987.tb13551.x","article-title":"Amino-acid sequence of human alpha 2-antiplasmin","volume":"166","author":"Lijnen","year":"1987","journal-title":"Eur. J. Biochem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1080\/15216540500404093","article-title":"The extraordinary ligand binding properties of human serum albumin","volume":"57","author":"Fasano","year":"2005","journal-title":"IUBMB Life"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1163","DOI":"10.1111\/j.1399-6576.2005.00733.x","article-title":"Colloids decrease clot propagation and strength: Role of factor XIII-fibrin polymer and thrombin-fibrinogen interactions","volume":"49","author":"Nielsen","year":"2005","journal-title":"Acta Anaesthesiol. Scand."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"700","DOI":"10.7326\/0003-4819-95-6-700","article-title":"Coagulopathy associated with hematin treatment for acute intermittent porphyria","volume":"95","author":"Morris","year":"1981","journal-title":"Ann. Intern. Med."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1182\/blood.V61.2.243.243","article-title":"Hematin: Unique effects of hemostasis","volume":"61","author":"Glueck","year":"1983","journal-title":"Blood"},{"key":"ref_34","first-page":"99","article-title":"The fibrinolysis system: Regulation of activity and physiologic functions of its main components","volume":"67","author":"Dobrovolsky","year":"2002","journal-title":"Biochemistry"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"28163","DOI":"10.1074\/jbc.C112.360800","article-title":"Human serum albumin can regulate amyloid-\u03b2 peptide fiber growth in the brain interstitium: Implications for Alzheimer disease","volume":"287","author":"Stanyon","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2570S","DOI":"10.1093\/jn\/nxaa079","article-title":"Histidine Metabolism and Function","volume":"150","author":"Brosnan","year":"2020","journal-title":"J. Nutr."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"3005","DOI":"10.1016\/j.jtha.2023.08.014","article-title":"Fibrinogen and fibrin: Synthesis, structure, and function in health and disease","volume":"21","author":"Wolberg","year":"2023","journal-title":"J. Thromb. Haemost."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2881","DOI":"10.1182\/blood.2020008951","article-title":"Plasminogen: An enigmatic zymogen","volume":"137","author":"Keragala","year":"2021","journal-title":"Blood"},{"key":"ref_39","first-page":"191","article-title":"\u03b12-Antiplasmin: New Insights and Opportunities for Ischemic Stroke","volume":"43","author":"Reed","year":"2017","journal-title":"Semin. Thromb. Hemost."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/8\/3567\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:12:13Z","timestamp":1760029933000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/8\/3567"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,10]]},"references-count":39,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2025,4]]}},"alternative-id":["ijms26083567"],"URL":"https:\/\/doi.org\/10.3390\/ijms26083567","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,10]]}}}