Three Dimensional Structure of Human Fibrinogen under Aqueous Conditions Visualized by Atomic Force Microscopy

Marchant, Roger; Barb, Matthew D.; Shainoff, John R.; Eppell, Steven J.; Wilson, David L.; Siedlecki, Christopher A.

doi:10.1055/s-0038-1656109

Cited by 73 publications

(90 citation statements)

References 15 publications

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“…This appearance was in good agreement with prior images generated of fixed, dehydrated fibrinogen by transmission electron microscopy (Hall and Slayter, 1959). Similar AFM imaging of fibrinogen has been reported by Marchant and co-workers (Marchant et al, 1997;Sit and Marchant, 1999).…”

Section: Afm and Haemostatic Proteinssupporting

confidence: 91%

Atomic Force Microscopy in the Study of Macromolecular Interactions in Hemostasis and Thrombosis: Utility for Investigation of the Antiphospholipid Syndrome

Montigny

Quinn

et al. 2005

Force Microscopy

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Section: Afm and Haemostatic Proteinssupporting

confidence: 91%

Atomic Force Microscopy in the Study of Macromolecular Interactions in Hemostasis and Thrombosis: Utility for Investigation of the Antiphospholipid Syndrome

Montigny

Quinn

et al. 2005

Force Microscopy

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“…Our in situ results for fibrinogen adsorption onto mica were similar to those observed by Ta et al 30 We were unable to reproduce the work of that group or of several others on HOPG and other hydrophobic surfaces, in situ. [32][33][34] We were only able to produce acceptable images of fibrinogen on the HOPG surface with our ex situ method. This is likely attributable to the effects of water hydration on the nanomechanical properties of the fibrinogen in air and solution.…”

mentioning

confidence: 97%

“…19,34 Additionally, the footprint of the molecule may be different on different surfaces. 19,33,34 Because observations were made only of area fraction coverage and not number or mass of protein attached, our kinetics and isotherm analysis only reflect area fraction coverage regardless of the mechanism of coverage (e.g., adsorption vs spreading). As noted in the past, however, perhaps the simple Langmuir analysis is not a sufficient model for the adsorption phenomenon.…”

mentioning

confidence: 99%

Quantification of the kinetics and thermodynamics of protein adsorption using atomic force microscopy

Gettens

Bai

Gilbert

2005

J Biomedical Materials Res

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Both in situ and ex situ methods for quantifying area fraction coverage of protein on a surface using atomic force microscopy were developed. The in situ method used a continuous fluid flow system to observe the kinetics of adsorption in real time. The ex situ method required immersing the sample in solution, drying the sample, and imaging in an ambient environment to obtain kinetic and isothermal data. These methods were developed using the plasma protein fibrinogen in a phosphate-buffered saline solution on grade IV muscovite mica and highly ordered pyrolytic graphite (HOPG) substrates. Kinetic and quasiisothermal data were obtained and a Langmuir model was fit to the data. An adsorption rate constant of 2.2 x 10(-4) mL . microg(-1)s(-1) and a desorption rate constant of 8.3 x 10(-5) s(-1) were found on an HOPG surface. Completely irreversible adsorption was found on the mica surface with an adsorption rate constant of 2.7 x 10(-4) mL . microg(-1)s(-1). Additionally, protein conformation and assembly orientation on these surfaces were documented where fibrinogen on HOPG formed a network-like structure, whereas fibrinogen on mica was more random. Also, nano-topographical factors (ledges) were seen as sites of preferential adsorption.

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“…1,2 The molecule has a symmetrical trinodal structure with a central E domain connected to 2 peripheral D domains by triple helix spacer arms. 3,4 After proteolytic activation by thrombin, fibrin monomers spontaneously polymerize through D:E interactions to form a half staggered bimolecular array that is stabilized by noncovalent end-to-end D:D interactions between adjacent monomer units. 1,2 Covalent cross-links are subsequently inserted to buttress D:D interaction and form an insoluble clot, with the C terminal of the ␥ chain intimately involved in all of these interactions.…”

mentioning

confidence: 99%

Novel fibrinogen γ375 Arg→Trp mutation (fibrinogen aguadilla) causes hepatic endoplasmic reticulum storage and hypofibrinogenemia

et al. 2002

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The proposita and her sister had chronically elevated liver function test results, and needle biopsy specimens showed scattered eosinophilic inclusions within the hepatocytes. On immunoperoxidase staining, the inclusions reacted strongly with anti-fibrinogen antisera; on electron-microscopic (EM) examination, the material appeared confined to the endoplasmic reticulum (ER) and was densely packed into tubular structures with a swirling fingerprint appearance. Coagulation investigations showed low functional and antigenic fibrinogen concentrations that were indicative of hypofibrinogenemia. Amplification and DNA sequencing showed a heterozygous CGG3 TGG mutation at codon 375 of the fibrinogen ␥ chain gene. This novel ␥375 Arg3 Trp substitution segregated with hypofibrinogenemia in 3 family members and was absent from 50 normal controls. When purified plasma fibrinogen chains were examined by sodium dodecyl sulfate/polyacrylamide gel electrophoresis, reverse-phase chromatography, electrospray ionization mass spectrometry, and isoelectric focusing, only normal ␥ chains were detected. In conclusion, we propose that this nonconservative mutation causes a conformational change in newly synthesized molecules and that this provokes aggregation within the ER and in turn causes the observed hypofibrinogenemia. Whereas the mutation site, ␥375, is located in the ␥D domain at the jaws of the primary E-to-D polymerization site, purified plasma fibrinogen showed normal polymerization, supporting our contention that molecules with variant chains never reach the circulation but accumulate in the ER. ( Plasma fibrinogen is synthesized in liver hepatocytes, and a signal peptide is cotranslationally cleaved from each chain as it translocates into the rough endoplasmic reticulum (ER). Chain assembly begins at this point with the formation of A␣-␥ and B␤-␥ intermediates, 5 and the acquisition of an additional B␤ or A␣ chain respectively leads to the formation of an [A␣-B␤-␥] half molecule. 6 Dimerization and disulfide bonding completes the assembly process, and further modification in the Golgi network leads to maturation of Nlinked oligosaccharides at positions B␤ 364 and ␥ 52 and the hydroxylation, sulfation, and phosphorylation of other specific side chains. 1 In the Golgi secretory vesicles, a C-terminal propeptide is removed from the A␣ Abbreviations: ER, endoplasmic reticulum; EM, electron microscopy; PBS, phosphate-buffered saline.From the

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Three Dimensional Structure of Human Fibrinogen under Aqueous Conditions Visualized by Atomic Force Microscopy

Cited by 73 publications

References 15 publications

Atomic Force Microscopy in the Study of Macromolecular Interactions in Hemostasis and Thrombosis: Utility for Investigation of the Antiphospholipid Syndrome

Atomic Force Microscopy in the Study of Macromolecular Interactions in Hemostasis and Thrombosis: Utility for Investigation of the Antiphospholipid Syndrome

Quantification of the kinetics and thermodynamics of protein adsorption using atomic force microscopy

Novel fibrinogen γ375 Arg→Trp mutation (fibrinogen aguadilla) causes hepatic endoplasmic reticulum storage and hypofibrinogenemia

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