Changes in adsorbed fibrinogen upon conversion to fibrin.
The conversion of adsorbed fibrinogen to fibrin in the presence of the enzyme thrombin was studied using surface plasmon resonance (SPR), a quartz crystal microbalance (QCM), sum frequency generation (SFG), atomic force microscopy (AFM), and an elutability assay. Exposure of adsorbed fibrinogen to thrombin resulted in a mass loss at the surface consistent with fibrinopeptide release and conversion to fibrin. Changes in hydration upon conversion of adsorbed fibrinogen to fibrin were determined from comparisons of acoustic (QCM) and optical (SPR) mass adsorption data. Conversion to fibrin also resulted in the adsorbed layer becoming more strongly bound to the surface and more compact. The elutability of adsorbed fibrinogen by Triton X-100, studied with SPR, decreased from 90 +/- 5 to 6 +/- 2% after conversion to fibrin. The height of the adsorbed monolayer, as determined by AFM, decreased from 5.5 +/- 2.2 to 1.7 +/- 0.8 nm. We conclude that thrombin-catalyzed fibrinopeptide release triggers significant changes in fibrinogen conformation beyond peptide cleavage.
Evans-Nguyen
Kenyon M
KM
Department of Chemistry, University of North Carolina at Chapel Hill, North Carolina 27599, USA.
Fuierer
Ryan R
RR
Fitchett
Brian D
BD
Tolles
Lauren R
LR
Conboy
John C
JC
Schoenfisch
Mark H
MH
D000268:Adhesiveness; D000327:Adsorption; D005337:Fibrin / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D005340:Fibrinogen / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D018625:Microscopy, Atomic Force; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D020349:Surface Plasmon Resonance; D013499:Surface Properties; D013917:Thrombin / Q000378:metabolism
D010446:Peptide Fragments; D005337:Fibrin; D005340:Fibrinogen; D013917:Thrombin