FTIR investigation of the secondary structure of type I collagen: New insight into the amide III band

2020 ◽  
Vol 229 ◽  
pp. 118006 ◽  
Author(s):  
Chiaramaria Stani ◽  
Lisa Vaccari ◽  
Elisa Mitri ◽  
Giovanni Birarda
Keyword(s):  
Secondary Structure ◽  
Type I Collagen ◽  
Type I ◽  
Insight Into ◽  
Ftir Investigation

scholarly journals Characterization of porcine osteonectin extracted from foetal calvariae

1988 ◽  
Vol 253 (1) ◽  
pp. 139-151 ◽  
Author(s):  
C Domenicucci ◽  
H A Goldberg ◽  
T Hofmann ◽  
D Isenman ◽  
S Wasi ◽  
...  
Keyword(s):  
Amino Acids ◽  
Secondary Structure ◽  
Type I Collagen ◽  
Culture Shock ◽  
Gel Filtration ◽  
Alpha Helix ◽  
Type I ◽  
Homologous Proteins ◽  
Compact Structure ◽  
Significant Difference

Osteonectin, extracted from foetal porcine calvariae with 0.5 M-EDTA, was purified to homogeneity by using gel filtration and polyanion anion-exchange fast protein liquid chromatography under dissociative conditions without the need of reducing agents. The purified protein migrated with an Mr of 40,300 on SDS/polyacrylamide gels and was similar to bovine osteonectin in both amino acid composition and in its ability to bind to hydroxyapatite in the presence of 4 M-guanidinium hydrochloride (GdmCl). However, unlike the bovine protein, porcine osteonectin did not bind selectively to hydroxyapatite when EDTA tissue extracts were used. In addition, purified porcine osteonectin did not show any apparent affinity for either native or denatured type I collagen, but did bind to serum albumin. Primary sequence analysis revealed an N-terminal alanine residue, with approximately one-half of the subsequent 35 residues identified as small hydrophobic amino acids and one-quarter as acidic amino acids. The only significant difference between the N-terminal sequences of the bovine and porcine proteins was the deletion of the tripeptide Val-Ala-Glu in porcine osteonectin. In contrast with bovine osteonectin, far-u.v.c.d. of porcine osteonectin revealed considerable secondary structure, of which 27% was alpha-helix and 39% was beta-sheet. Cleavage of the molecule with CNBr under non-reducing conditions generated five fragments, of which two major fragments (Mr 27,900 and 12,400) stained blue with Stains All, a reagent that stains sialic-acid-rich proteins/phosphate-containing proteins and/or Ca2+-binding proteins blue while staining other proteins pink. The 12,400-Mr fragment bound 45Ca2+ selectively, indicating a Ca2+-binding site in this part of the molecule. The 27,900-Mr fragment did not bind Ca2+, and since biosynthetic studies with 32PO4(3-) did not show phosphorylation of porcine osteonectin, this fragment is likely to be highly acidic. The incomplete cleavage of the molecule with CNBr and the ability of the molecule to regain its secondary structure after exposure to 7 M-urea are features consistent with the molecule having a compact structure that is stabilized by numerous disulphide bridges. The chemical and binding properties of porcine osteonectin are closely similar to the recently described ‘culture shock’, SPARC and BM-40 proteins, indicating that these are homologous proteins.

Type I collagen gene regulation and the molecular pathogenesis of cirrhosis

1993 ◽  
Vol 264 (4) ◽  
pp. G589-G595 ◽  
Author(s):  
D. A. Brenner ◽  
J. Westwick ◽  
M. Breindl
Keyword(s):  
Protein Interactions ◽  
Type I Collagen ◽  
Collagen Type ◽  
Collagen Type I ◽  
Molecular Pathogenesis ◽  
Matrix Proteins ◽  
Type I ◽  
And Function ◽  
Collagen Genes ◽  
Insight Into

Cirrhosis is characterized by an increased deposition of extracellular matrix proteins, including type I collagen. Type I collagen is a product of two genes, alpha 1(I) and alpha 2(I), which are generally coordinately regulated. Since expression of type I collagen genes is increased during cirrhosis, understanding the structure and function of the regulatory components of the type I collagen genes should provide insight into the molecular pathogenesis of cirrhosis. This review will analyze the collagen alpha 1(I) gene with respect to chromatin structure, DNA methylation, regulation by agonists, and DNA-protein interactions.

scholarly journals Synergist ablation induces rapid tendon growth through the synthesis of a neotendon matrix

2014 ◽  
Vol 117 (11) ◽  
pp. 1287-1291 ◽  
Author(s):  
Jonathan P. Gumucio ◽  
Anthony C. Phan ◽  
David G. Ruehlmann ◽  
Andrew C. Noah ◽  
Christopher L. Mendias
Keyword(s):  
Mechanical Loading ◽  
Type I Collagen ◽  
Fluorescent Microscopy ◽  
Type I ◽  
Fibroblast Cells ◽  
Cellular Mechanisms ◽  
Fast Green ◽  
Cross Sectional ◽  
Plantaris Tendon ◽  
Insight Into

Mechanical loading can increase tendon cross-sectional area (CSA), but the mechanisms by which this occurs are largely unknown. To gain a greater understanding of the cellular mechanisms of adult tendon growth in response to mechanical loading, we used a synergist ablation model whereby a tenectomy of the Achilles tendon was performed to induce growth of the synergist plantaris tendon. We hypothesized that after synergist ablation progenitor cells in the epitenon would proliferate and increase the size of the existing tendon matrix. Adult male mice were subjected to a bilateral Achilles tenectomy, and plantaris tendons were isolated from mice at 0, 2, 7, 14, and 28 days after surgery. Tendons were sectioned stained with either fast green and hematoxylin, prepared for fluorescent microscopy, or prepared for gene expression of scleraxis and type I collagen. After overload, there was a dramatic increase in total CSA of tendons, whereas the size of the original tendon matrix was not changed. Growth primarily occurred through the formation of a neotendon matrix between the original tendon and the epitenon, and contained cells that were proliferative and scleraxis positive. Additionally, an initial expansion of fibroblast cells occurred before the synthesis of new extracellular matrix. Fibroblasts in the original tendon did not re-enter the cell cycle. The results from this study provide new insight into the mechanisms of tendon growth, indicate tendon consists mostly of postmitotic cells, and that growth of tendon primarily occurs from the most superficial layers outward.

Hydration Effects on the Viscoelastic Properties of Collagen

2005 ◽  
Vol 898 ◽  
Author(s):  
M Ntim ◽  
Amanpreet Bembey ◽  
Virginia Ferguson ◽  
Andrew Bushby
Keyword(s):  
Elastic Modulus ◽  
Dynamic Mechanical Analysis ◽  
Viscoelastic Properties ◽  
Type I Collagen ◽  
Mechanical Analysis ◽  
Hierarchical Organization ◽  
Type I ◽  
Dynamic Mechanical ◽  
Insight Into

AbstractThe manner in which liquid interacts with collagen is unclear, with changes in hydration presenting ambiguity. At present, elastic modulus values for collagen quoted range from MPa to GPa. Dynamic mechanical analysis (DMA) of collagen in isolation provides an insight into the mechanical changes due to altered hydration states.Changes in the viscoelastic properties of collagen were examined as the material was systematically dehydrated in a series of water:solvent mixes to examine effects of dehydration. The effect of solvents with varying polarity was also examined. Tails from 11-week old wild type mice were used. Mouse tail is a tissue with a well-defined, hierarchical organization of type I collagen. The viscoelastic response of collagen was measured using dynamic mechanical analysis (DMA) in fiber extension mode over the frequency range of 1Hz to 10Hz. Samples were sequentially dehydrated in a series of solvent concentrations: 70% ethanol to 100% ethanol to 100% acetone and 70% ethanol to 70% methanol to 100% methanol for at least 1h. Selectively removing and then replacing water from collagen samples provides insight into the role of water in the ultrastructure of the tissue from the corresponding changes in the experimentally determined elastic modulus and viscous energy.

Insight into the collagen-degrading activity of a serine protease in the latex of Ficus carica cultivar Masui Dauphine

2021 ◽  
Author(s):  
Kosaku Nishimura ◽  
Keisuke Higashiya ◽  
Naoki Ueshima ◽  
Kenji Kojima ◽  
Teisuke Takita ◽  
...  
Keyword(s):  
Serine Protease ◽  
Gel Electrophoresis ◽  
Type I Collagen ◽  
Ficus Carica ◽  
Collagenolytic Activity ◽  
Type I ◽  
Partial Amino Acid Sequence ◽  
Two Dimensional Gel Electrophoresis ◽  
Genome Database ◽  
Insight Into

Abstract Ficus carica produces, in addition to the cysteine protease ficin, a serine protease. Earlier study on a serine protease from F. carica cultivar Brown Turkey showed that it specifically degraded collagen. In this study, we characterized the collagenolytic activity of a serine protease in the latex of F. carica cultivar Masui Dauphine. The serine protease degraded denatured, but not undenatured, acid-solubilized type I collagen. It also degraded bovine serum albumin, while the collagenase from Clostridium histolyticum did not. These results indicated that the serine protease in Masui Dauphine is not collagen-specific. The protease was purified to homogeneity by two-dimensional gel electrophoresis, and its partial amino acid sequence was determined by liquid chromatography-MS/MS. BLAST searches against the Viridiplantae (green plants) genome database revealed that the serine protease was a subtilisin-like protease. Our results contrast with the results of the earlier study stating that the serine protease from F. carica is collagen-specific.

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