title: Differences between neonates and adults in carbohydrate sequences and reaction kinetics of plasmin and alpha(2)-antiplasmin.
abstract: This study investigates reaction kinetics by slow-binding kinetics methods of both adult and fetal plasmin (Types 1 and 2) with adult and fetal alpha(2)-antiplasmin. In addition, carbohydrate sequences of Fetal and Adult Plasminogen Types 1 and 2, as well as fetal and adult alpha(2)-antiplasmin, were determined by mass spectrometric analysis. All curves of plasmin-alpha(2)-antiplasmin interaction followed the same pattern, indicating reversible slow-binding inhibition with an initial loose complex and a following tight complex. Differences between fetal and adult plasmin reactions with alpha(2)-antiplasmin were predominantly due to the initial loose complex. Values for K(i initial) in the reaction with adult alpha(2)-antiplasmin were 1.5 and 1.6 nM for Fetal Plasmin Types 1 and 2, respectively; compared to 0.3 and 0.7 nM for the corresponding adult types. Increasing concentrations of tranexamic acid resulted in a continuous increase of K(i initial) until a plateau was reached which was similar for all plasmin types. Almost identical values could be obtained when fetal alpha(2)-antiplasmin was used instead of adult alpha(2)-antiplasmin. Mass spectrometric analyses of the glycans present on plasminogen revealed a higher level of truncated N-glycans on the fetal material compared to the adult. The O-glycans of fetal and adult plasminogen were closely similar and only minor differences were observed between N-glycans of fetal and adult alpha(2)-antiplasmin. In conclusion, both fetal plasmin isoforms are less inhibited by alpha(2)-antiplasmin compared to the adult plasmin variants. These findings are important for the understanding of the physiology of the fibrinolytic system in neonates and provide further evidence that differences in glycosylation could be associated with marked effects on protein function.
authors.affiliation: Division of Haematology, National Institute for Biological Standards and Control, South Mimms, Hertfordshire, UK.
authors.forename: Martin
authors.initials: M
authors.lastname: Ries
authors.forename: Richard L
authors.initials: RL
authors.lastname: Easton
authors.forename: Colin
authors.initials: C
authors.lastname: Longstaff
authors.forename: Martin
authors.initials: M
authors.lastname: Zenker
authors.forename: Howard R
authors.initials: HR
authors.lastname: Morris
authors.forename: Anne
authors.initials: A
authors.lastname: Dell
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Gaffney
mesh_terms: D000328:Adult; D000375:Aging / Q000378:metabolism*; D002240:Carbohydrate Sequence; D005341:Fibrinolysin / Q000378:metabolism*; D006801:Humans; D007231:Infant, Newborn; D007700:Kinetics; D013058:Mass Spectrometry; D000979:alpha-2-Antiplasmin / Q000378:metabolism*
chemical_list: D000979:alpha-2-Antiplasmin; D005341:Fibrinolysin