scholarly journals Characterization of sequence specific binding of LARP6 to the 5’ stem-loop of type I collagen mRNAs and implications for rational design of antifibrotic drugs

2020 ◽  
Author(s):  
Lela Stefanovic ◽  
Blaine H. Gordon ◽  
Robert Silvers ◽  
Branko Stefanovic
Keyword(s):  
Rational Design ◽  
Type I Collagen ◽  
Specific Binding ◽  
High Rate ◽  
Stable Complex ◽  
Type I ◽  
Stem Loop ◽  
Rrm Domain ◽  
Α Helix ◽  
Antifibrotic Drugs

AbstractExcessive synthesis of type I collagen characterizes fibrotic diseases. Binding of LARP6 to the 5’ stem-loop (5’SL) of collagen mRNAs regulates their translation and the high rate of biosynthesis in fibrosis. LARP6 needs two domains to form stable complex with 5’SL RNA, the La-domain and the juxtaposed RRM domain (jointly called the La-module). We describe that the La-domain of LARP6 is necessary and sufficient for recognition of 5’SL in sequence specific manner. The three amino acid motif, RNK, located in the flexible loop which connects the second α-helix to the β-sheet of the La domain is critical for binding. Mutation of any of these three amino acids abolishes the binding of La-domain to 5’SL. The major site of crosslinking of LARP6 to 5’SL RNA was mapped to this motif. The RNK motif is not found in other LARPs, which can not bind 5’SL. Presence of RRM increases the stability of complex between La-domain and 5’SL RNA and RRM domain does not make extensive contacts with 5’SL RNA. We propose a model in which the initial recognition of 5’SL by LARP6 is mediated by the RNK epitope and further stabilized by the RRM domain. This discovery suggests that the interaction between LARP6 and collagen mRNAs can be blocked by small molecules that target the RNK epitope and will help rational design of the LARP6 binding inhibitors as specific antifibrotic drugs.

Mutation of the 5' untranslated region stem-loop mRNA structure reduces type I collagen deposition and arterial stiffness in male obese mice

2021 ◽  
Author(s):  
Francisco I. Ramirez-Perez ◽  
Makenzie L. Woodford ◽  
Mariana Morales-Quinones ◽  
Zachary I. Grunewald ◽  
Francisco J Cabral-Amador ◽  
...  
Keyword(s):  
Insulin Resistance ◽  
Type 2 Diabetes ◽  
Arterial Stiffness ◽  
Type I Collagen ◽  
Type I ◽  
Collagen Deposition ◽  
Wild Type ◽  
Stem Loop ◽  
Arterial Stiffening

Arterial stiffening, a characteristic feature of obesity and type 2 diabetes, contributes to the development and progression of cardiovascular diseases (CVD). Currently, no effective prophylaxis or therapeutics is available to prevent or treat arterial stiffening. A better understanding of the molecular mechanisms underlying arterial stiffening is vital to identify newer targets and strategies to reduce CVD burden. A major contributor to arterial stiffening is increased collagen deposition. In the 5' untranslated regions of mRNAs encoding for type I collagen, an evolutionally conserved stem-loop (SL) structure plays an essential role in its stability and post-transcriptional regulation. Here, we show that feeding a high fat/high sucrose (HFHS) diet for 28 weeks increases adiposity, insulin resistance, and blood pressure in male wild-type littermates. Moreover, arterial stiffness, assessed in vivo via aortic pulse wave velocity, and ex vivo using atomic force microscopy in aortic explants or pressure myography in isolated femoral and mesenteric arteries, was also increased in those mice. Notably, all these indices of arterial stiffness, along with collagen type I levels in the vasculature, were reduced in HFHS-fed mice harboring a mutation in the 5'SL structure, relative to wild-type littermates. This protective vascular phenotype in 5'SL-mutant mice did not associate with a reduction in insulin resistance or blood pressure. These findings implicate the 5'SL structure as a putative therapeutic target to prevent or reverse arterial stiffening and CVD associated with obesity and type 2 diabetes.

‘Small’-proteoglycan: collagen interactions: Keratan sulphate proteoglycan associates with rabbit corneal collagen fibrils at the ‘a’ and ‘c’ bands

1985 ◽  
Vol 5 (9) ◽  
pp. 765-774 ◽  
Author(s):  
J. E. Scott ◽  
M. Haigh
Keyword(s):  
Binding Sites ◽  
Type I Collagen ◽  
Specific Binding ◽  
Corneal Stroma ◽  
Collagen Fibrils ◽  
Type I ◽  
Keratan Sulphate ◽  
Protein Rich ◽  
Small Proteoglycan ◽  
Corneal Collagen

l. Proteoglycans (PGs) in rabbit corneal stroma and mouse sclera have been stained for electron microscopy with Cupromeronic blue in a critical electrolyte concentration (CEC) mode, with and without prior digestion of the tissue by keratanase or chondroitinase ABC to remove the keratan sulphate (KS) or chondroitin-dermatan sulphates (CS or DS) respectively.2. Two classes of PGs, located orthogonally to the corneal collagen fibrils at either the ‘step’ (band ‘a’ or ‘c’) or gap zone (band ‘d’ or ‘e’) are shown to be KS-PGs or DS-PGs respectively. Four separate and specific PG binding sites on Type I collagen fibrils have thus been identified.3. Rabbit corneal KS and DS PGs each contain two kinds of PG (Gregory JD, Coster L & Damle SP (1982) J. Biol. Chem.257, 6965–6970). We propose that each ‘small’ protein-rich PG is associated with a specific binding site on the collagen fibril.

scholarly journals Characterization of binding of LARP6 to the 5’ stem-loop of collagen mRNAs: Implications for synthesis of type I collagen

RNA Biology ◽  
2014 ◽  
Vol 11 (11) ◽  
pp. 1386-1401 ◽  
Author(s):  
Lela Stefanovic ◽  
Liam Longo ◽  
Yujie Zhang ◽  
Branko Stefanovic
Keyword(s):  
Type I Collagen ◽  
Type I ◽  
Stem Loop

Type I collagen shows a specific binding affinity for bovine dentin phosphophoryn

1986 ◽  
Vol 38 (3) ◽  
pp. 135-141 ◽  
Author(s):  
W. G. Stetler-Stevenson ◽  
Arthur Veis
Keyword(s):  
Binding Affinity ◽  
Type I Collagen ◽  
Specific Binding ◽  
Type I

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 Characterization of Acid-Soluble Collagen from Food Processing By-Products of Snakehead Fish (Channa striata)

Processes ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 1188
Author(s):  
Thi Mong Thu Truong ◽  
Van Muoi Nguyen ◽  
Thanh Truc Tran ◽  
Thi Minh Thuy Le
Keyword(s):  
Acid Content ◽  
Type I Collagen ◽  
Type I ◽  
Fish Skin ◽  
Soluble Collagen ◽  
Acid Soluble Collagen ◽  
Imino Acid ◽  
Channa Striata ◽  
Α Helix ◽  
By Products

The isolation of acid-soluble collagen (ASC) from by-products of snakehead fish (Channa striata), including skin and the mixture of skin and scale, has been investigated. The recovery yield of fish skin ASC (13.6%) was higher than ASC from fish skin and scale (12.09%). Both ASCs were identified as type I collagen and showed maximal solubility at pH 2. Collagen samples from the mixture of skin and scale had higher imino acid content (226 residues/1000 residues) and lower wavenumber in the amide I and amide III region (1642 and 1203 cm−1, respectively) than the fish skin ASC (the imino acid content was 220 residues/1000 residues and the wavenumber in the amide I and amide III were 1663 and 1206 cm−1, respectively. The difference scanning calorimeter (DSC) showed higher thermal stability in ASC from the mixture of skin and scale (Td of 35.78 °C) than fish skin ASC (34.21 °C). From the result, the denaturation temperature of ASC had a close relationship with the content of imino acid as well as with the degradation of α-helix in amide I and III. These results suggest that collagen could be obtained effectively from snakehead fish by-products and has potential as a realistic alternative to mammalian collagens.

scholarly journals Three-dimensional printed hydroxyapatite bone tissue engineering scaffold with antibacterial and osteogenic ability

2021 ◽  
Vol 15 (1) ◽  
Author(s):  
Liu Zhongxing ◽  
Wu Shaohong ◽  
Li Jinlong ◽  
Zhang Limin ◽  
Wang Yuanzheng ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Bone Defect ◽  
Type I Collagen ◽  
Porous Scaffold ◽  
Specific Binding ◽  
Three Dimensional ◽  
Protein Translation ◽  
Inhibition Zone ◽  
Bone Morphogenetic Protein 2 ◽  
Type I

AbstractThe development of an effective scaffold for bone defect repair is an urgent clinical need. However, it is challenging to design a scaffold with efficient osteoinduction and antimicrobial activity for regeneration of bone defect. In this study, we successfully prepared a hydroxyapatite (HA) porous scaffold with a surface-specific binding of peptides during osteoinduction and antimicrobial activity using a three-dimensional (3D) printing technology. The HA binding domain (HABD) was introduced to the C-terminal of bone morphogenetic protein 2 mimetic peptide (BMP2-MP) and antimicrobial peptide of PSI10. The binding capability results showed that BMP2-MP and PSI10-containing HABD were firmly bound to the surface of HA scaffolds. After BMP2-MP and PSI10 were bound to the scaffold surface, no negative effect was observed on cell proliferation and adhesion. The gene expression and protein translation levels of type I collagen (COL-I), osteocalcin (OCN) and Runx2 have been significantly improved in the BMP2-MP/HABP group. The level of alkaline phosphatase significantly increased in the BMP2-MP/HABP group. The inhibition zone test against Staphylococcus aureus and Escherichia coli BL21 prove that the PSI10/HABP@HA scaffold has strong antibacterial ability than another group. These findings suggest that 3D-printed HA scaffolds with efficient osteoinduction and antimicrobial activity represent a promising biomaterial for bone defect reconstruction.

scholarly journals Binding of LARP6 to the Conserved 5′ Stem–Loop Regulates Translation of mRNAs Encoding Type I Collagen

2010 ◽  
Vol 395 (2) ◽  
pp. 309-326 ◽  
Author(s):  
Le Cai ◽  
Dillon Fritz ◽  
Lela Stefanovic ◽  
Branko Stefanovic
Keyword(s):  
Type I Collagen ◽  
Type I ◽  
Stem Loop
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