scholarly journals Identification of the sites in collagen α-chains that bind serum anti-gelatin factor (cold-insoluble globulin)

1978 ◽  
Vol 169 (1) ◽  
pp. 55-59 ◽  
Author(s):  
W Dessau ◽  
B C Adelmann ◽  
R Timpl
Keyword(s):  
Active Region ◽  
Guinea Pig ◽  
Affinity Chromatography ◽  
Human Serum ◽  
Type I Collagen ◽  
Type I ◽  
Inhibition Assay ◽  
Serum Factor ◽  
Cold Insoluble Globulin

Anti-gelatin factor was prepared from guinea-pig and human serum by affinity chromatography on denatured type-I collagen. As shown previously, this component is related to cold-insoluble globulin. It reacted with 125I-labelled denatured collagen, and the reaction could be inhibited by preincubation with unlabelled collagenous components. In the inhibition assay comparable activities were observed for native and denatured type-I, -II, -III and -IV collagens. There was also no difference in reactivity between collagens of different species. The reactive sites in the collagen alpha-chains were located by inhibition assays on distinct CNBr- and collagenase-derived peptides. The results obtained with fragments from alpha1(I)-, alpha2- and alpha1(II)-chains indicate that the most active region is located between positions 643 and 819 of the alpha1-chain. Lower activities were found for other regions of collagen and may indicate that the factor has the potential to interact with several sites in the alpha-chains. The present data agree with observations by Kleinman, McGoodwin & Klebe [Biochem. Biophys. Res. Commun. (1976) 72, 426-432] on the specificity of a serum factor promoting the attachment of fibroblasts to collagen.

Phospholipase A2 activity is increased in guinea pig uterine cervix in late pregnancy and at parturition

1995 ◽  
Vol 269 (5) ◽  
pp. E940-E947 ◽  
Author(s):  
M. R. Rajabi ◽  
A. V. Cybulsky
Keyword(s):  
Guinea Pig ◽  
Type I Collagen ◽  
Late Pregnancy ◽  
Cervical Dilatation ◽  
Type I ◽  
Pla2 Activity ◽  
Phospholipases A2 ◽  
Rate Limiting Step ◽  
Cytosolic Pla2 ◽  
Rate Limiting

Release of arachidonic acid from phospholipids by phospholipases A2 (PLA2) is the rate-limiting step in prostaglandin (PG) synthesis. PGE2 and PGF2 alpha are essential intermediates in interstitial collagenase-mediated degradation of type I collagen, a key step in cervical dilatation at parturition. We demonstrate that PLA2 is present in cytosolic fractions of guinea pig cervix and PLA2 activity is increased during cervical dilatation at parturition. In the cervix of nonpregnant animals, PLA2 activity against phosphatidylcholine (PC) and phosphatidylethanolamine (PE) was 13 +/- 6 and 49 +/- 27 pmol.min-1.mg protein-1, respectively. Levels were similar at day 25 of pregnancy. At day 50, PLA2-PC increased to 190 +/- 54 pmol.min-1.mg-1, and PLA2-PE rose to 592 +/- 127 pmol.min-1.mg-1. At parturition (68 +/- 2 days) there were further increases of 49-67%. PLA2 activity declined toward basal levels 2 days postpartum. Almost 50% of the enhanced cytosolic PLA2 (cPLA2) activity at day 50 or at parturition was of high molecular mass and was identified as the "85-kDa cPLA2." Increases in cPLA2 activity at these time points were not, however, associated with increases in cPLA2 protein or phosphorylation of cPLA2, compared with nonpregnant animals. This study suggests that multiple PLA2 enzymes, including cPLA2, are involved in cervical dilatation at parturition.

Isolation And Purification Of Collagen And α1 Receptor From Platelet Membrane

1981 ◽  
Author(s):  
T M Chiang ◽  
A H Kang
Keyword(s):  
Affinity Chromatography ◽  
Binding Site ◽  
Gel Electrophoresis ◽  
Type I Collagen ◽  
Specific Binding ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Type I ◽  
Isolation And Purification ◽  
Platelet Membranes

We have previously demonstrated that chick skin type I collagen and the α1(I) chain mediate platelet aggregation. Aggregation is associated with specific binding of these substances by platelet membranes. We now describe the isolation and purification of the receptor. Platelet membranes were prepared as described previously and isolated membranes were solubilized in 0.5% Triton. The receptor was then purified by a combination of gel filtration, affinity chromatography on α1(I)-sepharose or type I collagen-sepharose and preparative polyacrylamide gel electrophoresis. The receptor activity was assayed either directly by a binding assay using (14C)-glycine-labeled α1(I) or indirectly by an adhesion inhibition assay on Sepharose 2B with (14C)-sero- tinin-labeled platelets.The results show that the α1(I) receptor can be purified to a single band on SDS-gel electrophoresis with a recovery of 2.5%. Its activity is destroyed by preincubation with trypsin or pronase indicating it is a protein. The apparent molecular weight as estimated by gel filtration and SDS-gel electrophoresis is 95,000 daltons. The binding of (14C)- labeled α1(I) is specifically displaced by unlabeled α1(I), and the bound radioactivity can be removed by treatment with purified bacterial collagenase. The binding of (14C)- labeled α1(I) by the purified α1(I) receptor can also be inhibited by the receptor isolated from collagen-sepharose affinity chromatography. These data suggest that the α1(I) binding site is identical to the collagen binding site.

scholarly journals Expression of Wnt/β-Catenin Signaling Pathway and Its Regulatory Role in Type I Collagen with TGF-β1 in Scleral Fibroblasts from an Experimentally Induced Myopia Guinea Pig Model

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Min Li ◽  
Ying Yuan ◽  
Qingzhong Chen ◽  
Rao Me ◽  
Qing Gu ◽  
...  
Keyword(s):  
Guinea Pig ◽  
Signaling Pathway ◽  
Type I Collagen ◽  
Intervention Group ◽  
Neutralizing Antibody ◽  
Self Control ◽  
Pig Model ◽  
Type I ◽  
Pathway Expression ◽  
Guinea Pig Model

Background. To investigate Wnt/β-catenin signaling pathway expression and its regulation of type I collagen by TGF-β1 in scleral fibroblasts from form-deprivation myopia (FDM) guinea pig model.Methods. Wnt isoforms were examined using genome microarrays. Scleral fibroblasts from FDM group and self-control (SC) group were cultured. Wnt isoforms,β-catenin, TGF-β1, and type I collagen expression levels were examined in the two groups with or without DKK-1 or TGF-β1 neutralizing antibody.Results. For genome microarrays, the expression of Wnt3 in FDM group was significantly greater as confirmed in retinal and scleral tissue. The expression of Wnt3 andβ-catenin significantly increased in FDM group and decreased significantly with DKK-1. TGF-β1 expression level decreased significantly in FDM group and increased significantly with DKK-1. Along with morphological misalignment inside and outside cells, the amount of type I collagen decreased in FDM group. Furthermore, type I collagen increased and became regular in DKK-1 intervention group, whereas it decreased and rearranged more disorder in TGF-β1 neutralizing antibody intervention group.Conclusions. The activation of Wnt3/β-catenin signaling pathway was demonstrated in primary scleral fibroblasts in FDM. This pathway further reduced the expression of type I collagen by TGF-β1, which ultimately played a role in scleral remodeling during myopia development.

Aerosolized Polymerized Type I Collagen Reduces Airway Inflammation and Remodelling in a Guinea Pig Model of Allergic Asthma

Lung ◽  
2009 ◽  
Vol 188 (2) ◽  
pp. 97-105 ◽  
Author(s):  
Paola Moreno-Alvarez ◽  
Edgar Sánchez-Guerrero ◽  
Erasmo Martínez-Cordero ◽  
Rogelio Hernández-Pando ◽  
María G. Campos ◽  
...  
Keyword(s):  
Guinea Pig ◽  
Airway Inflammation ◽  
Allergic Asthma ◽  
Type I Collagen ◽  
Pig Model ◽  
Type I ◽  
Guinea Pig Model

Regulation of pancreatic duct epithelial growth in vitro

1990 ◽  
Vol 258 (6) ◽  
pp. G833-G840 ◽  
Author(s):  
T. B. Verme ◽  
S. R. Hootman
Keyword(s):  
Guinea Pig ◽  
Pancreatic Duct ◽  
Type I Collagen ◽  
Basal Medium ◽  
Trophic Factors ◽  
Type I ◽  
Trophic Effect ◽  
Initial Area

Effects of a number of possible trophic factors on growth of guinea pig pancreatic duct epithelial monolayers were investigated. Isolated fragments of main and interlobular ducts were prepared and explanted onto both tissue culture plastic and thick gels of type I collagen. Monolayers growing out from explants were first cultured in a basal medium for 3 or 4 days. Next, the medium was supplemented individually with bombesin, carbachol, caerulein, epidermal growth factor (EGF), secretin, 12-O-tetradecanoylphorbol 13-acetate (TPA), or vasoactive intestinal peptide (VIP). Cells were cultured in the absence or presence of these possible trophic factors, and monolayer areas were determined morphometrically at 0, 2, and 4 days. Rate of growth was determined from increase in area over each 2-day period. Monolayers grown in basal medium alone on plastic increased to 479% of initial area over the 4-day test period; those grown on collagen increased to 523%. Explants cultured in presence of bombesin, carbachol, caerulein, secretin, TPA, and VIP on either substrate grew at rates not significantly different from those cultured in basal medium. By contrast, duct monolayers grown on plastic or collagen in presence of 10 nM EGF expanded in area to 722 and 1,070%, respectively, of their initial areas. The EC50 for this trophic effect was approximately 1 nM. These results show that EGF exerts a potent trophic effect on guinea pig pancreatic duct cells in vitro but also indicate that cell division in the pancreatic main and interlobular ducts is not regulated by caerulein and related peptide hormones that have been reported to have growth-promoting effects on exocrine pancreas in vivo.

scholarly journals Mechanism of Human Dermal Fibroblast Migration Driven by Type I Collagen and Platelet-derived Growth Factor-BB

2004 ◽  
Vol 15 (1) ◽  
pp. 294-309 ◽  
Author(s):  
Wei Li ◽  
Jianhua Fan ◽  
Mei Chen ◽  
Shengxi Guan ◽  
David Sawcer ◽  
...  
Keyword(s):  
Growth Factor ◽  
Human Serum ◽  
Type I Collagen ◽  
Collagen Matrix ◽  
Gene Cassette ◽  
Platelet Derived Growth Factor ◽  
Type I ◽  
Kinase Gene ◽  
Extracellular Signal Regulated Kinase ◽  
Extracellular Signal

Migration of human dermal fibroblasts (HDFs) is critical for skin wound healing. The mechanism remains unclear. We report here that platelet-derived growth factor-BB (PDGF-BB) is the major promotility factor in human serum for HDF motility on type I collagen. PDGF-BB recapitulates the full promotility activity of human serum and anti-PDGF neutralizing antibodies completely block it. Although collagen matrix initiates HDF migration without growth factors, PDGF-BB–stimulated migration depends upon attachment of the cells to a collagen matrix. The PDGF-BB's role is to provide directionality and further enhancement for the collagen-initiated HDF motility. To study the collagen and PDGF-BB “dual signaling” in primary HDF, we establish “gene cassettes” plus lentiviral gene delivery approach, in which groups of genes are studied individually or in combination for their roles in HDF migration. Focal adhesion kinase, p21Rac,CDC42-activated kinase and Akt are grouped into an upstream kinase gene cassette, and the four major mitogen-activated protein kinases (extracellular signal-regulated kinase 1/2, p38, c-Jun NH2-terminal kinase, and extracellular signal-regulated kinase 5) are grouped into a downstream kinase gene cassette. The experiments demonstrate 1) the genes' individual roles and specificities, 2) their combined effects and sufficiency, and 3) the mechanisms of their intermolecular connections in HDF migration driven by collagen and PDGF-BB.

Development of an assay for the quantification of type I collagen synthesis in the guinea pig

2005 ◽  
Vol 297 (1-2) ◽  
pp. 133-141 ◽  
Author(s):  
Helen L. Quasnichka ◽  
John F. Tarlton ◽  
Janet M. Anderson-MacKenzie ◽  
Michael E.J. Billingham ◽  
Allen J. Bailey ◽  
...  
Keyword(s):  
Guinea Pig ◽  
Collagen Synthesis ◽  
Type I Collagen ◽  
Type I

Collagen fibrillogenesis in vitro: interaction of types I and V collagen

1986 ◽  
Vol 44 ◽  
pp. 210-211
Author(s):  
Arthur J. Wasserman ◽  
Kathy C. Kloos ◽  
David E. Birk
Keyword(s):  
Type I Collagen ◽  
Corneal Stroma ◽  
Minor Component ◽  
Homogeneous Distribution ◽  
Type I ◽  
Type V Collagen ◽  
Fibril Morphology ◽  
Type V ◽  
In Vitro Interaction

Type I collagen is the predominant collagen in the cornea with type V collagen being a quantitatively minor component. However, the content of type V collagen (10-20%) in the cornea is high when compared to other tissues containing predominantly type I collagen. The corneal stroma has a homogeneous distribution of these two collagens, however, immunochemical localization of type V collagen requires the disruption of type I collagen structure. This indicates that these collagens may be arranged as heterpolymeric fibrils. This arrangement may be responsible for the control of fibril diameter necessary for corneal transparency. The purpose of this work is to study the in vitro assembly of collagen type V and to determine whether the interactions of these collagens influence fibril morphology.

948: Clinical Safety, Histologic Findings and Surgical Methods in Complex Phalloplasty Using Type I Collagen

2007 ◽  
Vol 177 (4S) ◽  
pp. 314-314 ◽  
Author(s):  
Joon-Yang Kim ◽  
Hoon Seog Jean ◽  
Beom Joon Kim ◽  
Kye Yong Song
Keyword(s):  
Type I Collagen ◽  
Type I ◽  
Clinical Safety ◽  
Surgical Methods
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