Differences Between Neonates and Adults in Tissue-Type-Plasminogen Activator (t-PA)-Catalyzed Plasminogen Activation With Various Effectors and in Carbohydrate Sequences of Fibrinogen Chains

• Published in: Thrombosis research 2001-08, Vol.103 (3), p.173-184
• Main Authors: Ries, M., Easton, R.L., Longstaff, C., Zenker, M., Corran, P.H., Morris, H.R., Dell, A., Gaffney, P.J.
• Format: Article
• Language: English
• Subjects: Activation kinetics; Adult; Carbohydrate Sequence; Enzyme Activation - drug effects; Fibrin; Fibrin - pharmacology; Fibrinogen - chemistry; Fibrinogen - pharmacology; Gas Chromatography-Mass Spectrometry; Glycosylation; Humans; Infant, Newborn; Kinetics; Neonates; Peptide Fragments - chemistry; Peptide Fragments - pharmacology; Plasminogen; Plasminogen - metabolism; Spectrometry, Mass, Fast Atom Bombardment; Tissue Plasminogen Activator - metabolism
• ISSN: 0049-3848; 1879-2472
• DOI: 10.1016/S0049-3848(01)00289-4

Our study investigates the effect of fetal and adult soluble fibrin (SF), fetal and adult fibrinogen Aα- and γ-chains, as well as adult CNBr-fibrinogen fragments on tissue-type plasminogen activator (t-PA)-catalyzed plasminogen activation of both fetal and adult Glu-plasminogen types 1 and 2. In addition, we determined carbohydrate sequences of fetal and adult Bβ- and γ-chains by mass spectrometric analysis. In the absence of an effector, no substantial differences in the rate of plasmin formation could be seen between the fetal and adult plasminogen types. In the presence of an effector, both fetal Glu-plasminogen types revealed lower values for k cat app than the respective adult types. No differences could be seen in the values for K m app. The resulting differences in catalytic efficiencies between the fetal and adult plasminogen types were much less than previously reported. No differences could be seen between fetal and adult effectors in stimulating t-PA-catalyzed plasminogen activation. Detailed analyses of the activation kinetics revealed a longer initial phase of slow plasmin formation of both fetal Glu-plasminogen types compared to their respective adult types, indicating a slower plasmin-induced modification of CNBr-fibrinogen fragments or SF by fetal plasmin. Mass spectrometric analysis of the N-glycans present on adult and fetal Bβ- and γ-fibrinogen chains showed the presence of a major monosialylated biantennary structure with lesser amounts of the disialylated form. In contrast to previous data, we conclude that catalytic efficiency of t-PA-catalyzed plasminogen activation in neonates is only slightly lower than in adults.