The role of ascorbate and histidine in fibrinogen protection against changes following exposure to a sterilizing dose of gamma-irradiation.
Sodium ascorbate and histidine were employed to protect fibrinogen against modifications followed by a gamma-irradiation process that could potentially inactivate the blood-borne viruses in plasma-derived products. Fibrinogen was irradiated (50 kGy total dose, on dry ice) using a 60Co source. Samples were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot. Carbonyl groups were measured by the 2,4-dinitrophenylhydrazine-coupled method, and the fibrinogen clotting activity was assessed by different functional assays. In irradiated fibrinogen, the carbonyl group concentration was elevated three-fold versus control; and moderate fragmentation of largely Aalpha and Bbeta chains was revealed. The rate of thrombin-catalyzed fibrinogen polymerization was inhibited (average 50%) with normal fibrinopeptide release and with a minor decrease of total clottable fibrinogen and alpha-polymer formation. Ascorbate reduced the incorporation of carbonyls to the fibrinogen molecule (by > 50% at 50 mmol/l; P < 0.001). Contrary to ascorbate, which alone delayed the fibrinogen polymerization rate, histidine abolished irradiation-induced inhibition of fibrinogen polymerization (by 80% at 50 mmol/l; P < 0.001). In conclusion, even though ascorbate effectively protects fibrinogen from oxidation due to its adverse effects on fibrinogen function, it may not serve as a suitable radioprotective. On the contrary, the first definite evidence is provided that radiation-sterilized fibrinogen in the presence of histidine greatly retains its clotting capability.
Zbikowska
Halina Malgorzata
HM
Department of General Biochemistry, University of Lodz, Lodz, Poland. zbikow@biol.uni.lodz.pl
Nowak
Pawel
P
Wachowicz
Barbara
B
D001205:Ascorbic Acid / Q000737:chemistry / Q000494:pharmacology / Q000502:physiology*; D001688:Biological Products; D005176:Factor XIII / Q000378:metabolism / Q000528:radiation effects; D005337:Fibrin / Q000378:metabolism / Q000528:radiation effects; D005340:Fibrinogen / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects*; D005344:Fibrinopeptide A / Q000378:metabolism; D005345:Fibrinopeptide B / Q000378:metabolism; D005720:Gamma Rays; D006639:Histidine / Q000737:chemistry / Q000494:pharmacology / Q000502:physiology*; D006801:Humans; D066298:In Vitro Techniques; D050050:Protein Carbonylation / Q000528:radiation effects; D011837:Radiation-Protective Agents / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D013242:Sterilization / Q000379:methods; D013917:Thrombin / Q000378:metabolism; D013918:Thrombin Time / Q000379:methods
D001688:Biological Products; D011837:Radiation-Protective Agents; D005344:Fibrinopeptide A; D005345:Fibrinopeptide B; D006639:Histidine; D005337:Fibrin; D005340:Fibrinogen; D005176:Factor XIII; D013917:Thrombin; D001205:Ascorbic Acid