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Fibrinogen is a heme-associated, carbon monoxide sensing molecule: a preliminary report
Journal article   Peer reviewed

Fibrinogen is a heme-associated, carbon monoxide sensing molecule: a preliminary report

Vance G. Nielsen, Jack B. Cohen, S. Nini Malayaman, Mathew Nowak and Keith Vosseller
Blood coagulation & fibrinolysis, v 22(5), pp 443-447
01 Jul 2011
PMID: 21451399

Abstract

Hematology Life Sciences & Biomedicine Science & Technology
The objective of this study was to determine how carbon monoxide directly modifies fibrinogen utilizing liquid chromatography-mass spectrometry (LC-MS/MS) by examining fibrinogen exposed to carbon monoxide releasing molecule-2 [tricarbonyldichlororuthenium (II) dimer; CORM-2]. Purified fibrinogen was exposed to 0, 25, 50 or 100 mu mol/l CORM-2 for 5 min at 37 degrees C and then stored at -80 degrees C before analyses with LC-MS/MS. In a second series of experiments, normal plasma was exposed to 0 or 100 mu mol/l CORM-2 in the absence or presence of the nitric oxide donor sodium nitroprusside and hydroquinone (an organic reductant) to compete with carbon monoxide binding to a putative heme group found on fibrinogen. Coagulation was activated with tissue factor (n=8 per condition). Thrombus growth was monitored with thrombelastography for 15 min. LC-MS/MS did not detect any direct modifications of amino acids in fibrinogen, but detection of small regions of both the alpha and gamma chains was lost following exposure to CORM-2 and endoproteinase digestion with trypsin and Glu-C. An ion with the same m/z and expected retention time as heme was found in the purified fibrinogen. Exposure of plasma to nitric oxide/hydroquinone significantly decreased CORM-2-mediated enhancement of coagulation without affecting the coagulation kinetics of plasma not exposed to CORM-2. Carbon monoxide derived from CORM-2 likely modifies fibrinogen via modulation of a fibrinogen-associated heme group(s). Whereas the precise molecular location of heme attachment and three-dimensional conformational change secondary to carbon monoxide exposure remain to be determined, fibrinogen appears to be a carbon monoxide sensing molecule. Blood Coagul Fibrinolysis 22:443-447 (C) 2011 Wolters Kluwer Health vertical bar Lippincott Williams & Wilkins.

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