In Situ Single-Molecule AFM Investigation of Surface-Induced Fibrinogen Unfolding on Graphite

Dubrovin, Evgeniy V.; Barinov, Nikolay A.; Schäffer, Tilman E.; Klinov, Dmitry V.

doi:10.1021/acs.langmuir.9b01178

Cited by 13 publications

(32 citation statements)

References 44 publications

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“…The observed transformation of fibrinogen and fibrinogen–myeloperoxidase complexes from trinodular (Figure 1a) to predominantly fibrillary (Figure 1d) structures can be associated with fibrinogen unfolding (denaturation), which is induced by a fibrinogen contact with myeloperoxidase molecules. Similar unfolding of fibrinogen conformation has been observed after several minutes of fibrinogen adsorption on a GM-HOPG surface 26,27 . However, in the present work the contribution of surface-induced unfolding of fibrinogen molecules is negligible due to relatively small time of adsorption (5 s) 26–29 .…”

Section: Resultssupporting

confidence: 74%

“…Similar unfolding of fibrinogen conformation has been observed after several minutes of fibrinogen adsorption on a GM-HOPG surface 26,27 . However, in the present work the contribution of surface-induced unfolding of fibrinogen molecules is negligible due to relatively small time of adsorption (5 s) [26][27][28][29] .…”

Section: Afm Characterization Of Fibrinogen Unfolding By Myeloperoxidasesupporting

confidence: 74%

“…Three types of trinodular structures can be distinguished depending on a height of the outer globules (see the height profiles in Figure 1b along the dotted lines in the enlarged regions of AFM images): the structures with the outer globules 3 -3.5 nm in height (e.g., region III) can be assigned to fibrinogen molecules, which haven't formed complexes with myeloperoxidase, whereas the structures with significantly higher one (e.g., region I) or both (e.g., region II) outer globulesto the fibrinogen-myeloperoxidase complexes with 1:1 or 1:2 fibrinogen-myeloperoxidase molar ratio, respectively. The height distribution of the globules of trinodular structures (Figure 1c) demonstrates the peaks around 2 and 3.2 nm that correspond to inner and outer globules of native-like fibrinogen molecules (e.g., region III) respectively [27][28][29] . Moreover, it has one more maximum at ~4 nm that we associate with fibrinogen-myeloperoxidase complexes.…”

Section: Afm Characterization Of Fibrinogen Unfolding By Myeloperoxidasementioning

confidence: 99%

“…Fibrinogen molecules adsorbed onto a GM-HOPG surface for 5 s are characterized by a trinodular structure with two outer globular regions 3 -3.5 nm high and one central globular region ~2 nm high [27][28][29] . Myeloperoxidase molecules deposited by the same procedure adsorb onto a GM-HOPG surface as globular structures ~4 nm high 30 .…”

Section: Afm Characterization Of Fibrinogen Unfolding By Myeloperoxidasementioning

confidence: 99%

“…Recently we have visualized unfolding of individual fibrinogen molecules induced by their contact with a highly oriented graphite (HOPG) surface modified with N,N’-(decane-1,10-diyl)bis(tetraglycineamide) (GM) ([Gly 4 -NHCH 2 ] 2 C 10 H 20 ) 26,27 . We have also developed an approach of AFM characterization of unfolding of individual protein molecules induced by the factors other than contact with a substrate surface.…”

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Myeloperoxidase-induced fibrinogen unfolding and clotting

Barinov¹,

Pavlova

Tolstova

et al. 2021

Preprint

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Fibrinogen is a major protein of blood coagulation system and is a promising component of biomaterials and protein matrixes. Conformational changes of fibrinogen underlie the important mechanism of thrombin mediated fibrinogen clotting but also may induce the loss of its biological activity and (amyloid) aggregation. Understanding and controlling of fibrinogen unfolding is important for the development of fibrinogen based materials with tunable properties. We have discovered that myeloperoxidase induces denaturation of fibrinogen molecules followed by fibrinogen clotting, which is not thrombin-dependent. This is the first example of ATP-independent, non-targeted protein-induced protein denaturation. The morphological structure of unfolded fibrinogen molecules and “non-conventional” fibrinogen clots has been characterized using high-resolution atomic force microscopy and scanning electron microscopy techniques. Circular dichroism (CD) spectroscopy has shown no significant changes of the secondary structure of the fibrinogen clots. The absorbance spectrophotometry has demonstrated that the kinetics of myeloperoxidase induced fibrinogen clotting strongly decays with growth of ionic strength indicating a major role of the Debye screening effect in regulating of this process. The obtained results provide with the novel concepts of protein unfolding and open new insights into fibrinogen clotting. Moreover, they give new possibilities in biotechnological and biomedical applications, e.g., for regulation of fibrinogen clotting and platelet adhesion and for the development of fibrinogen-based matrices.The organization of a protein molecule is characterized by different hierarchical levels such as primary, secondary, tertiary and quaternary structure. Protein unfolding or denaturation, i.e. its transformation to a lower order structure (and loss of a higher order structure), is a biologically and biotechnologically relevant process. Protein unfolding is a prerequisite for an alternative folding pathway including amyloid aggregation 1,2. Unfolded proteins may be used in development of protein films and coatings with special properties such as enhanced mechanical stability 3–5, resistance to protein adsorption or platelet adhesion 6,7 and other advantages 8. Unfolding of a protein molecule may lead to the loss of its biological function 9 that has important consequences in biosensor 10,11 and pharmaceutical applications 12.

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Section: Resultssupporting

confidence: 74%

Section: Afm Characterization Of Fibrinogen Unfolding By Myeloperoxidasesupporting

confidence: 74%

Section: Afm Characterization Of Fibrinogen Unfolding By Myeloperoxidasementioning

confidence: 99%

Section: Afm Characterization Of Fibrinogen Unfolding By Myeloperoxidasementioning

confidence: 99%

mentioning

confidence: 99%

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Myeloperoxidase-induced fibrinogen unfolding and clotting

Barinov¹,

Pavlova

Tolstova

et al. 2021

Preprint

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Myeloperoxidase‐induced fibrinogen unfolding and clotting

Barinov

Pavlova

Tolstova

et al. 2022

Microscopy Res & Technique

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Principles of Fibrinogen Fiber Assembly In Vitro

Stamboroski

Joshi

Noeske

et al. 2021

Macromolecular Bioscience

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Fibrinogen nanofibers hold great potential for applications in wound healing and personalized regenerative medicine due to their ability to mimic the native blood clot architecture. Although versatile strategies exist to induce fibrillogenesis of fibrinogen in vitro, little is known about the underlying mechanisms and the associated length scales. Therefore, in this manuscript the current state of research on fibrinogen fibrillogenesis in vitro is reviewed. For the first time, the manifold factors leading to the assembly of fibrinogen molecules into fibers are categorized considering three main groups: substrate interactions, denaturing and non‐denaturing buffer conditions. Based on the meta‐analysis in the review it is concluded that the assembly of fibrinogen is driven by several mechanisms across different length scales. In these processes, certain buffer conditions, in particular the presence of salts, play a predominant role during fibrinogen self‐assembly compared to the surface chemistry of the substrate material. Yet, to tailor fibrous fibrinogen scaffolds with defined structure–function‐relationships for future tissue engineering applications, it still needs to be understood which particular role each of these factors plays during fiber assembly. Therefore, the future combination of experimental and simulation studies is proposed to understand the intermolecular interactions of fibrinogen, which induce the assembly of soluble fibrinogen into solid fibers.

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In Situ Single-Molecule AFM Investigation of Surface-Induced Fibrinogen Unfolding on Graphite

Cited by 13 publications

References 44 publications

Myeloperoxidase-induced fibrinogen unfolding and clotting

Myeloperoxidase-induced fibrinogen unfolding and clotting

Myeloperoxidase‐induced fibrinogen unfolding and clotting

Principles of Fibrinogen Fiber Assembly In Vitro

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