title: Conformation of human fibrinogen in solution from polarized triplet spectroscopy.
abstract: The rotational motions of human fibrinogen in solution at 20 degrees C have been examined, in the 0.2-12-microseconds time range, by measuring the laser-induced dichroism of the triplet state of an erythrosin probe covalently bonded to the protein. The decay of the anisotropy was multiexponential, and up to three correlation times (phi 1 = 380 +/- 50 ns, phi 2 = 1.1 +/- 0.1 microseconds, and phi 3 = 3.3 +/- 0.6 microseconds) were needed to obtain a satisfactory analysis. The experimental data are consistent with the brownian motions of an elongated, rigid particle. If the correlation times are combined with previous data on the intrinsic viscosity of fibrinogen, the rotational and translational diffusive properties of the protein can be reproduced with high accuracy by idealizing it as an elongated ellipsoid of revolution with dimensions (2a x 2b) of (54 +/- 6) x (7.2 +/- 0.5) nm, having rotational diffusion constants of D parallel = (6.2 +/- 0.7) x 10(5) s-1 and D perpendicular = (5 +/- 1) x 10(4) s-1. The possibility of Ca(2+)-dependent changes in the rigidity or conformation of fibrinogen was excluded by examining the submicrosecond time-resolved fluorescence depolarization of 1-methylpyrene conjugates of the protein in the presence of different calcium concentrations. Although there are inherent difficulties to extrapolate the data on isolated fibrinogen molecules to the polymerizing species, this relatively stiff conformation meets the requirements of the classical half-staggered double-stranded model of fibrin polymerization rather better than those of the recently proposed interlocked single-stranded mechanism.
authors.affiliation: Instituto de Química Física, CSIC, Serrano, Madrid, Spain.
authors.forename: J M
authors.initials: JM
authors.lastname: Montejo
authors.forename: K R
authors.initials: KR
authors.lastname: Naqvi
authors.forename: M P
authors.initials: MP
authors.lastname: Lillo
authors.forename: J
authors.initials: J
authors.lastname: González-Rodríguez
authors.forename: A U
authors.initials: AU
authors.lastname: Acuña
mesh_terms: D002942:Circular Dichroism; D004923:Erythrosine; D005340:Fibrinogen / Q000737:chemistry* / Q000648:ultrastructure; D005454:Fluorescence Polarization; D006801:Humans; D008433:Mathematics; D008854:Microscopy, Electron; D008962:Models, Theoretical; D011485:Protein Binding; D011487:Protein Conformation; D011721:Pyrenes; D012996:Solutions
chemical_list: D011721:Pyrenes; D012996:Solutions; D005340:Fibrinogen; C053035:1-methylpyrene; D004923:Erythrosine