Collagen Structure and Stability

Shoulders, Matthew D.; Raines, Ronald T.

doi:10.1146/annurev.biochem.77.032207.120833

Cited by 3,015 publications

(2,720 citation statements)

References 155 publications

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“…46 In the case of organofunctional group, the chlorine (Cl) present in CPTES has a hydrophobic character, whereas the epoxy group present in GPTES is more hydrophilic due to the presence of an oxygen that can establish hydrogen bonds with water at the surface. Moreover, silanized GPTES surfaces resulted in a lower amount of silane molecules than silanized CPTES surfaces as shown by XPS measurements (Table 2 and This finding is consistent with the presence of hydrocarbon chains and hydrophilic functional groups in the collagen molecules, 47 and with the fact that underlying silanes can be partially exposed on the surface. …”

Section: Xpssupporting

confidence: 81%

Different Organization of Type I Collagen Immobilized on Silanized and Nonsilanized Titanium Surfaces Affects Fibroblast Adhesion and Fibronectin Secretion

Marín-Pareja

Cantini

González-García

et al. 2015

ACS Appl. Mater. Interfaces

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ABSTRACT:Silanization has emerged in recent years as a way to obtain a stronger and more stable attachment of biomolecules to metallic substrates. However, its impact on protein conformation, a key aspect that influences cell response, has hardly been studied. In this work we analyzed by AFM the distribution and conformation of type I collagen on plasma treated surfaces before and after silanization. Subsequently, we investigated the effect of the different collagen conformations on fibroblasts adhesion and fibronectin secretion by immunofluorescence analyses. Two different organosilanes were used on plasma-treated titanium surfaces, either 3-chloropropyl-triethoxy-silane (CPTES), or 3-Glycidyloxypropyl-triethoxy-silane (GPTES). The properties and amount of the adsorbed collagen was assessed by contact angle, XPS, OWLS and AFM.AFM studies revealed different conformations of type I collagen depending on the silane employed. Collagen was organized in fibrillar networks over very hydrophilic (plasma treated titanium) or hydrophobic (silanized with CPTES) surfaces, the latter forming little globules with a beads-on-a-string appearance, whereas over surfaces presenting an intermediate hydrophobic character (silanized with GPTES), collagen was organized into clusters with a size increasing at higher protein concentration in solution.Cell response was strongly affected by collagen conformation, especially at low collagen density. The samples exhibiting collagen organized in globular clusters (GPTES-functionalized samples) favored a faster and better fibroblast adhesion, as well as better cell spreading, focal adhesions formation and more pronounced fibronectin fibrillogenesis. In contrast, when a certain protein concentration was reached at the material surface the effect of collagen conformation was masked, and similar fibroblast response was observed in all samples.

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Section: Xpssupporting

confidence: 81%

Different Organization of Type I Collagen Immobilized on Silanized and Nonsilanized Titanium Surfaces Affects Fibroblast Adhesion and Fibronectin Secretion

Marín-Pareja

Cantini

González-García

et al. 2015

ACS Appl. Mater. Interfaces

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“…All collagens consist of a set of three polypeptide chains held together in a helical conformation via hydrogen bonding. Each polypeptide chain is characterized by a repeating Gly‐X‐Y sequence where X corresponds frequently to proline and Y to hydroxyproline (Shoulders & Raines 2009; Nguyen et al . 2012).…”

Section: Discussionmentioning

confidence: 99%

Label‐free Raman spectroscopic imaging to extract morphological and chemical information from a formalin‐fixed, paraffin‐embedded rat colon tissue section

Gaifulina

Maher

Kendall

et al. 2016

Int J Experimental Path

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SummaryAnimal models and archived human biobank tissues are useful resources for research in disease development, diagnostics and therapeutics. For the preservation of microscopic anatomical features and to facilitate long‐term storage, a majority of tissue samples are denatured by the chemical treatments required for fixation, paraffin embedding and subsequent deparaffinization. These aggressive chemical processes are thought to modify the biochemical composition of the sample and potentially compromise reliable spectroscopic examination useful for the diagnosis or biomarking. As a result, spectroscopy is often conducted on fresh/frozen samples. In this study, we provide an extensive characterization of the biochemical signals remaining in processed samples (formalin fixation and paraffin embedding, FFPE) and especially those originating from the anatomical layers of a healthy rat colon. The application of chemometric analytical methods (unsupervised and supervised) was shown to eliminate the need for tissue staining and easily revealed microscopic features consistent with goblet cells and the dense populations of cells within the mucosa, principally via strong nucleic acid signals. We were also able to identify the collagenous submucosa‐ and serosa‐ as well as the muscle‐associated signals from the muscular regions and blood vessels. Applying linear regression analysis to the data, we were able to corroborate this initial assignment of cell and tissue types by confirming the biological origin of each layer by reference to a subset of authentic biomolecular standards. Our results demonstrate the potential of using label‐free Raman microspectroscopy to obtain superior imaging contrast in FFPE sections when compared directly to conventional haematoxylin and eosin (H&E) staining.

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“…The formation of Hyp is essential to stabilize the collagen triple helix and confer its thermal stability at body temperature (Shoulders and Raines 2009). Lysyl hydroxylation is involved in the formation of covalent intra-and inter-molecular crosslinks, contributing to condensation and fibril formation (Takaluoma et al 2007).…”

Section: Introductionmentioning

confidence: 99%

Recombinant expression of hydroxylated human collagen in Escherichia coli

Rutschmann

Baumann

Cabalzar

et al. 2013

Appl Microbiol Biotechnol

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Collagen is the most abundant protein in the human body and thereby a structural protein of considerable biotechnological interest. The complex maturation process of collagen, including essential post-translational modifications such as prolyl and lysyl hydroxylation, has precluded large-scale production of recombinant collagen featuring the biophysical properties of endogenous collagen. The characterization of new prolyl and lysyl hydroxylase genes encoded by the giant virus mimivirus reveals a method for production of hydroxylated collagen. The coexpression of a human collagen type III construct together with mimivirus prolyl and lysyl hydroxylases in Escherichia coli yielded up to 90 mg of hydroxylated collagen per liter culture. The respective levels of prolyl and lysyl hydroxylation reaching 25 % and 26 % were similar to the hydroxylation levels of native human collagen type III. The distribution of hydroxyproline and hydroxylysine along recombinant collagen was also similar to that of native collagen as determined by mass spectrometric analysis of tryptic peptides. The triple helix signature of recombinant hydroxylated collagen was confirmed by circular dichroism, which also showed that hydroxylation increased the thermal stability of the recombinant collagen construct. Recombinant hydroxylated collagen produced in E. coli supported the growth of human umbilical endothelial cells, underlining the biocompatibility of the recombinant protein as extracellular matrix. The high yield of recombinant protein expression and the extensive level of prolyl and lysyl hydroxylation achieved indicate that recombinant hydroxylated collagen can be produced at large scale for biomaterials engineering in the context of biomedical applications.

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Collagen Structure and Stability

Cited by 3,015 publications

References 155 publications

Different Organization of Type I Collagen Immobilized on Silanized and Nonsilanized Titanium Surfaces Affects Fibroblast Adhesion and Fibronectin Secretion

Different Organization of Type I Collagen Immobilized on Silanized and Nonsilanized Titanium Surfaces Affects Fibroblast Adhesion and Fibronectin Secretion

Label‐free Raman spectroscopic imaging to extract morphological and chemical information from a formalin‐fixed, paraffin‐embedded rat colon tissue section

Recombinant expression of hydroxylated human collagen in Escherichia coli

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