title: Does topology drive fiber polymerization?
abstract: We have developed new procedures to examine the early steps in fibrin polymerization. First, we isolated fibrinogen monomers from plasma fibrinogen by gel filtration. Polymerization of fibrinogen monomers differed from that of plasma fibrinogen. The formation of protofibrils was slower and the transformation of protofibrils to fibers faster for the fibrinogen monomers. Second, we used formaldehyde to terminate the polymerization reactions. The formaldehyde-fixed products obtained at each time point were examined by dynamic light scattering and transmission electron microscopy (TEM). The data showed the formaldehyde-fixed products were stable and representative of the reaction intermediates. TEM images showed monomers, short oligomers, protofibrils, and thin fibers. The amount and length of these species varied with time. Short oligomers were less than 5% of the molecules at all times. Third, we developed models that recapitulate the TEM images. Fibrin monomer models were assembled into protofibrils, and protofibrils were assembled into two-strand fibers using Chimera software. Monomers were based on fibrinogen crystal structures, and the end-to-end interactions between monomers were based on D-dimer crystal structures. Protofibrils assembled from S-shaped monomers through asymmetric D:D interactions were ordered helical structures. Fibers were modeled by duplicating a protofibril and rotating the duplicate 120° around its long axis. No specific interactions were presumed. The two protofibrils simply twisted around one another to form a fiber. This model suggests that the conformation of the protofibril per se promotes the assembly into fibers. These findings introduce a novel mechanism for fibrin assembly that may be relevant to other biopolymers.
authors.affiliation: Department of Pathology and Laboratory Medicine and ‡Department of Computer Science, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina 27599, United States.
authors.forename: Lihong
authors.initials: L
authors.lastname: Huang
authors.forename: Joe Ping-Lin
authors.initials: JP
authors.lastname: Hsiao
authors.forename: Camilla
authors.initials: C
authors.lastname: Powierza
authors.forename: Russell M
authors.initials: RM
authors.lastname: Taylor
authors.forename: Susan T
authors.initials: ST
authors.lastname: Lord
mesh_terms: D000818:Animals; D001777:Blood Coagulation; D030562:Databases, Protein; D019281:Dimerization; D005337:Fibrin / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D005340:Fibrinogen / Q000737:chemistry / Q000378:metabolism; D005404:Fixatives / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D046529:Microscopy, Electron, Transmission; D008958:Models, Molecular; D008970:Molecular Weight; D009391:Nephelometry and Turbidimetry; D058105:Polymerization; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D059748:Proteolysis; D013499:Surface Properties; D013917:Thrombin / Q000378:metabolism
chemical_list: D005404:Fixatives; D005557:Formaldehyde; D005337:Fibrin; D005340:Fibrinogen; D013917:Thrombin