Bioactive hydrogels based on Designer Collagens

Cosgriff‐Hernandez, Elizabeth; Hahn, Mariah S.; Russell, Brooke; Wilems, Thomas S.; Muñoz-Pinto, Dany J.; Browning, Mary Beth; Rivera, José Julián; Höök, Magnus

doi:10.1016/j.actbio.2010.05.002

Cited by 94 publications

(181 citation statements)

References 31 publications

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“…Previous studies show the insertion of human collagen sequences within the recombinant triple-helical protein conferred biological activity similar to that found in human collagens. For example, recombinant Scl2.28 proteins with inserted integrin binding sequences (GFPGER, GFPGEN, and GLPGER) were shown to bind cell surface integrins in vitro and to mediate cell binding (23,39), whereas insertion of the GRPGKRGKQGQK sequence led to heparin binding (40). Incorporation of 6 triplets from the human collagen type III MMP cleavage site led to digestion by MMP-1 at the same site as in native type III collagen (29).…”

Section: Discussionmentioning

confidence: 99%

Definition of the Native and Denatured Type II Collagen Binding Site for Fibronectin Using a Recombinant Collagen System

Abbonante

Yiğit

et al. 2014

Journal of Biological Chemistry

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Background: Sequence requirements for triple-helical collagen to bind fibronectin are not fully understood. Results: Fibronectin affinity was conferred on a recombinant bacterial collagen by incorporating specific human collagen II sequences. Conclusion:The chimeric collagen defines the minimum collagen II sequence required for fibronectin affinity. Significance: This system is useful to investigate sequence/activity relationships for triple-helical collagen and to generate scalable bioactive materials.

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Section: Discussionmentioning

confidence: 99%

Definition of the Native and Denatured Type II Collagen Binding Site for Fibronectin Using a Recombinant Collagen System

Abbonante

Yiğit

et al. 2014

Journal of Biological Chemistry

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“…10 This protein can be readily prepared in good yields, up to around 19 g/L in 2 L fermentation studies 11 and can be purified using a simple precipitation and proteolysis approach. 12 The protein can be modified, either by specific sequence changes to introduce specific cell binding and other functional domains [12][13][14][15] or by chemical modifications, for example to produce stabilized and hydrogel materials and other functional domains. [12][13] These non-animal, bacterial collagens share the same characteristic triple-helical structure that is found in animal collagens and several other animal proteins.…”

Section: Introductionmentioning

confidence: 99%

“…12 The protein can be modified, either by specific sequence changes to introduce specific cell binding and other functional domains [12][13][14][15] or by chemical modifications, for example to produce stabilized and hydrogel materials and other functional domains. [12][13] These non-animal, bacterial collagens share the same characteristic triple-helical structure that is found in animal collagens and several other animal proteins. [16][17][18] However, the Scl2 collagen domain is small, about a quarter of the length, 234 residues, 19 of the abundant, mammalian interstitial collagens; eg type I, type II and type III collagens, which are just over 1000 residues each.…”

Section: Introductionmentioning

confidence: 99%

Preparation and characterization of monomers to tetramers of a collagen-like domain fromStreptococcus pyogenes

et al. 2014

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Abbreviations: V, non-collagenous N-terminal domain of the S. pyogenes collagen-like protein; CL, collagenous domain of the S.pyogenes collagen-like protein; VCL, full collagen-like protein structureThe collagen like domain Scl2 from Streptococcus pyogenes has been proposed as a potential biomedical material. It is non-cytotoxic and non-immunogenic and can be prepared in good yield in fermentation. The Scl2 collagen domain is about a quarter of the length, 234 residues, of the main collagen type, mammalian type I collagen (1014 residues) that is currently used in biomedical devices. In the present study we have made constructs comprising 1 to 4 copies of the Scl2 collagen domain, plus these same constructs with a CysCys sequence at the C-terminal, analogous to that found in mammalian type III collagens. The yields of these constructs were examined from 2 L fermentation studies. The yields of both series declined with increasing size. Circular dichroism showed that the addition of further collagen domains did not lead to a change in the melting temperature compared to the monomer domain. Addition of the CysCys sequence led to a small additional stabilization of about 2-3 C for the monomer construct when the folding (V) domain was present.

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“…Prokaryotic collagens also form higher order structures (14), as often observed with mammalian collagens (15). Because of these similarities to mammalian collagens, prokaryotic collagen-like proteins are a potential replacement of animal-derived collagens in medical and bioengineering applications (9,14,16,17).Integrins are cell surface heterodimeric receptors that participate in a variety of cellular processes through activation of different signaling pathways. There are four collagen-binding integrins, ␣1, ␣2, ␣10, and ␣11, which each complex with a ␤1 subunit to form a functioning receptor.…”

mentioning

confidence: 99%

An Engineered α1 Integrin-binding Collagenous Sequence

Seo

Russell

Rivera

et al. 2010

Journal of Biological Chemistry

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Collagen is an extracellular matrix structural component that can regulate cellular processes through its interaction with the integrins, ␣1␤1, ␣2␤1, ␣10␤1, and ␣11␤1. Collagen-like proteins have been identified in a number of bacterial species. Here, we used Scl2 from Streptococcus pyogenes serotype M28 strain MGAS6274 as a backbone for the introduction of discrete integrin-binding sequences. The introduced sequences GLPGER, GFPGER, or GFPGEN did not affect triple helix stability of the Scl (Streptococcal collagen-like) protein. Using ELISA and surface plasmon resonance, we determined that Scl2 GLPGER and Scl2 GFPGER bound to recombinant human ␣1 and ␣2 I-domains in a metal ion-dependent manner and without a requirement for hydroxyproline. We predicted a novel and selective integrinbinding sequence, GFPGEN, through the use of computer modeling and demonstrated that Scl2 GFPGEN shows specificity toward the ␣1 I-domain and does not bind the ␣2 I-domain. Using C2C12 cells, we determined that intact integrins interact with the modified Scl2 proteins with the same selectivity as recombinant I-domains. These modified Scl2 proteins also acted as cell attachment substrates for fibroblast, endothelial, and smooth muscle cells. However, the modified Scl2 proteins were unable to aggregate platelets. These results indicate that Scl2 is a suitable backbone for the introduction of mammalian integrin-binding sequences, and these sequences may be manipulated to individually target ␣1␤1 and ␣2␤1.Collagen is a major constituent of the extracellular matrix where it functions as a structural component. In addition, collagen can directly or indirectly interact with cellular receptors and regulate a variety of cellular processes (1). There are at least 28 identified mammalian collagens each consisting of three polypeptides that can be genetically identical or distinct (2). A defining feature of collagens is the tightly packed left-handed triple helix made of polypeptide segments with repeating GXY triplets. The small Gly residue fits in the interior of the triple helix, and the X and Y positions are often occupied by proline and hydroxyproline residues. Hydroxylation of proline residues stabilizes the triple helical structure and inhibition of posttranslational hydroxylation decreases the melting temperature of the mammalian collagen triple helix by ϳ15°C (3).Surface proteins with collagen-like domains recently have been found on a number of prokaryotic organisms, including Streptococcus pyogenes, Streptococcus equi, and Bacillus anthracis (4 -6). These collagen-like domains contain conserved GXY repeats but lack the hydroxyproline found in mammalian collagens. S. pyogenes often contain two collagen-like proteins, Scl1 and Scl2 (4, 7-13). The primary sequences and length of different domains, including the GXY repeat segments, vary considerably in Scl1 and Scl2 proteins from different strains (4). In electron micrographs, both proteins form a lollipop-like structure with an N-terminal variable globular domain connected to an extende...

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Bioactive hydrogels based on Designer Collagens

Cited by 94 publications

References 31 publications

Definition of the Native and Denatured Type II Collagen Binding Site for Fibronectin Using a Recombinant Collagen System

Definition of the Native and Denatured Type II Collagen Binding Site for Fibronectin Using a Recombinant Collagen System

Preparation and characterization of monomers to tetramers of a collagen-like domain fromStreptococcus pyogenes

An Engineered α1 Integrin-binding Collagenous Sequence

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