Screening for novel cell adhesive regions in bovine Achilles tendon collagen peptides

2014 ◽  
Vol 92 (1) ◽  
pp. 9-22 ◽  
Author(s):  
Pradipta Banerjee ◽  
Alka Mehta ◽  
C. Shanthi
Keyword(s):  
Cell Adhesion ◽  
Docking Studies ◽  
Exchange Chromatography ◽  
Adhesion Assay ◽  
Structural Protein ◽  
Cell Adherence ◽  
Collagen Peptides ◽  
Adhesion Ability ◽  
A Cell ◽  
Hydrophilic Residues

Collagen, a major structural protein of the ECM, is known for its high cell adherence capacity. This study was conducted to identify regions in collagen that harbour such bioactivity. Collagen from tendon was hydrolysed and the peptides fractionated using ion-exchange chromatography (IEC). Isolated peptide fractions were coated onto disposable dishes and screened for cell adherence and proliferative abilities. Active IEC fractions were further purified by chromatography, and two peptides, C2 and E1 with cell adhesion ability, were isolated. A cell adhesion assay done with different amounts of C2 coated onto disposable dishes revealed the maximum adhesion to be 94.6%, compared with 80% for collagen coated dishes and an optimum peptide coating density of 0.507 nmoles per cm2 area of the dish. Growth of cells on C2, collagen, and E1 revealed a similar pattern and a reduction in the doubling time compared with cells grown on uncoated dishes. C2 had a mass of 2.046 kDa with 22 residues, and sequence analysis revealed a higher percentage occurrence of hydrophilic residues compared with other regions in collagen. Docking studies revealed GDDGEA in C2 as the probable site of interaction with integrins α2β1 and α1β1, and stability studies proved C2 to be mostly protease-resistant.

Specthophotmetric quantification of cell-cell adherence by an enzyme-linked immuno-cell adhesion assay

1990 ◽  
Vol 132 (1) ◽  
pp. 51-56 ◽  
Author(s):  
Ineke Bruynzeel ◽  
Liesbeth M.H. Van der Raaij ◽  
Dick M. Boorsma ◽  
Peter De Haan ◽  
Rik J. Scheper ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Adhesion Assay ◽  
Cell Adherence ◽  
Cell Adhesion Assay ◽  
Cell Cell

Pentamidine Is a Specific, Non-Peptide, GPIIb/llla Antagonist

1996 ◽  
Vol 75 (03) ◽  
pp. 503-509 ◽  
Author(s):  
Dermot Cox ◽  
Toshiaki Aoki ◽  
Jiro Seki ◽  
Yukio Motoyama ◽  
Keizo Yoshida
Keyword(s):  
Monoclonal Antibody ◽  
Cell Adhesion ◽  
Endothelial Cell ◽  
Intracellular Ph ◽  
Platelet Adhesion ◽  
Adhesion Assay ◽  
Agonist Peptide ◽  
A Cell ◽  
Granule Release ◽  
Endothelial Cell Adhesion

SummaryPentamidine was previously shown to act on glycoprotein (GP) Ilb/IIIa (Cox et al., Thromb Haemost 1992; 68: 731). In this paper we study the effect of pentamidine on other RGD-dependent receptors. In a cell adhesion assay, pentamidine was 500 times more potent than RGDS at inhibiting platelet adhesion to fibrinogen. While RGDS inhibited platelet adhesion to fibronectin, endothelial cell adhesion to vitronectin or fibronectin, 293 cell adhesion to vitronectin, IMR 32 cell adhesion to fibronectin and C32 cell adhesion to vitronectin; pentamidine failed to inhibit these interactions at doses as high as 1 mM. Resting platelets fixed in the presence of 1 mM RGDS had increased binding of fibrinogen, i.e., RGDS activated GPIMIIa, while pentamidine at 100 ΜM had no effect. Similarly, RGDS induced the binding of an anti-LIBS monoclonal antibody, while pentamidine had no effect. Pentamidine partially, but significantly, inhibited lysosome and a-granule release induced by the thrombin agonist peptide, while RGDS had no effect. Neither pentamidine nor RGDS affected ADP-induced Ca2+ influx. Pentamidine had no effect on ADP-induced intracellular pH changes while RGDS prevented the pH from returning to normal. Thus, pentamidine is a non-peptide GPIIb/IIIa antagonist that is non-activating and is specific for GPIIb/IIIa.

Efficient synthesis of novel RGD based peptides and the conjugation of the pyrazine moiety to their N-terminus

2019 ◽  
Vol 43 (6) ◽  
pp. 2702-2709
Author(s):  
Fatima Hamdan ◽  
Zahra Bigdeli ◽  
Saeed Balalaie ◽  
Norbert Sewald ◽  
Carmela Michalek
Keyword(s):  
Cell Adhesion ◽  
Adhesion Assay ◽  
Efficient Synthesis ◽  
Cell Adhesion Assay ◽  
N Terminus ◽  
A Cell

Novel RGD based peptides (RGDFAKLF and RGDNGRG) were designed and synthesized and were later coupled to the pyrazine moiety at the N-terminus. The IC50 values from the in vitro study of the target peptides using a cell adhesion assay indicated the essential impact of the existence of the pyrazine moiety. Meanwhile, peptide 4 exhibited the best IC50 value.

Synthesis and SAR of a Library of Cell-Permeable Biotin-R8ERY* Peptidomimetics Inhibiting α4β7 Integrin Mediated Adhesion of TK-1 Cells to MAdCAM-1-Fc

2012 ◽  
Vol 65 (9) ◽  
pp. 1349 ◽  
Author(s):  
Stefanie Papst ◽  
Anaïs F. M. Noisier ◽  
Margaret A. Brimble ◽  
Yi Yang ◽  
Yih-Chih Chan ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Disease Type ◽  
Adhesion Assay ◽  
Tyrosine Residue ◽  
Regulatory Domain ◽  
Lead Compound ◽  
Disease States ◽  
A Cell ◽  
Inflammatory Bowel

The α4β7 integrin is a well‐known target for the development of drugs against various inflammatory disease states including inflammatory bowel disease, type 1 diabetes, and multiple sclerosis. The β7 subunit contains the cell adhesion regulatory domain (CARD) motif YDRREY within its cytoplasmic domain, which is an effective peptide agent for inhibiting T-cell adhesion. The synthesis of a library of cell-permeable β7 integrin inhibitors based on the shortened biotin-R8ERY (R8 = (l-arginine)8) motif is reported, wherein the tyrosine residue has been modified. The synthesised peptidomimetics were evaluated in a cell adhesion assay and shown to inhibit Mn2+-activated adhesion of mouse TK-1 T-cells to mouse MAdCAM-1. Several analogues exhibited improved activity to that of the tyrosine-containing lead compound 1 (biotin-R8ERY). Specifically, analogues 4, 10, and 22 bearing a 4-chloro, a 4-nitro, and a 3,3-diphenyl substituent showed an increase in activity of approximately two-fold compared with that of the initial lead compound. The six most active compounds of the tested series had IC50’s between 25 and 50 μM.

A fluorometric approach to the quantitation of cell number with application to a cell adhesion assay

1988 ◽  
Vol 110 (1) ◽  
pp. 93-100 ◽  
Author(s):  
David M. Lewinsohn ◽  
Brian J. Nickoloff ◽  
Eugene C. Butcher
Keyword(s):  
Cell Adhesion ◽  
Cell Number ◽  
Adhesion Assay ◽  
Cell Adhesion Assay ◽  
A Cell

scholarly journals Biological Performance of Electrospun Polymer Fibres

Materials ◽  
2019 ◽  
Vol 12 (3) ◽  
pp. 363 ◽  
Author(s):  
Ivan Hall Barrientos ◽  
Graeme MacKenzie ◽  
Clive Wilson ◽  
Dimitrios Lamprou ◽  
Paul Coats
Keyword(s):  
Cell Proliferation ◽  
Cell Adhesion ◽  
Cell Viability ◽  
Acid Group ◽  
Adhesion Assay ◽  
Biological Responses ◽  
Polymeric Scaffolds ◽  
Biological Studies ◽  
A Cell ◽  
Biological Performance

The evaluation of biological responses to polymeric scaffolds are important, given that the ideal scaffold should be biocompatible, biodegradable, promote cell adhesion and aid cell proliferation. The primary goal of this research was to measure the biological responses of cells against various polymeric and collagen electrospun scaffolds (polycaprolactone (PCL) and polylactic acid (PLA) polymers: PCL–drug, PCL–collagen–drug, PLA–drug and PLA–collagen–drug); cell proliferation was measured with a cell adhesion assay and cell viability using 5-bromo-2′-deoxyuridine (BrdU) and resazurin assays. The results demonstrated that there is a distinct lack of growth of cells against any irgasan (IRG) loaded scaffolds and far greater adhesion of cells against levofloxacin (LEVO) loaded scaffolds. Fourteen-day studies revealed a significant increase in cell growth after a 7-day period. The addition of collagen in the formulations did not promote greater cell adhesion. Cell viability studies revealed the levels of IRG used in scaffolds were toxic to cells, with the concentration used 475 times higher than the EC50 value for IRG. It was concluded that the negatively charged carboxylic acid group found in LEVO is attracting positively charged fibronectin, which in turn is attracting the cell to adhere to the adsorbed proteins on the surface of the scaffold. Overall, the biological studies examined in this paper are valuable as preliminary data for potential further studies into more complex aspects of cell behaviour with polymeric scaffolds.

Smart Hybrid Mussel Adhesive Materials for Cell and Tissue Engineering

2008 ◽  
Vol 47-50 ◽  
pp. 861-864
Author(s):  
Dong Soo Hwang ◽  
Seong Hye Lim ◽  
Hyung Joon Cha
Keyword(s):  
Tissue Engineering ◽  
Cell Adhesion ◽  
Cell Attachment ◽  
Attachment Site ◽  
Peptide Sequence ◽  
Adhesion Properties ◽  
Adhesion Ability ◽  
A Cell ◽  
Mussel Adhesive ◽  
Adhesive Materials

Mussel foot protein is widely known for candidate of strong adhesive materials. Six times fp-1 decapeptide repeats and fp-5 are chosen among the various type of foot protein and fused into (MAP) fp-151. Through various cell-adhesion analyses, we previously demonstrated that fp- 151 has the potential to be used as a cell or tissue bioadhesive. In the present study, to improve the cell-adhesion properties of fp-151, it was designed a new cell-adhesive protein, fp-151-RGD, which is a fusion with the GRGDSP residues, a RGD peptide sequence that has previously been identified at the cell-attachment site of fibronectin, at the C-terminus of fp-151. Also, recombinant fp-151- RGD maintained the advantages associated with fp-151, such as a high production yield in Escherichia coli and simple purification, it showed superior spreading ability, which is important for cell proliferation under serum-free conditions, as well as better cell-adhesion ability compared with other commercially produced cell-adhesion materials such as poly-L-lysine (PLL) and the naturally extracted MAP mixture Cell-Tak. The excellent adhesion and spreading abilities of fp- 151-RGD might be due to the fact that it utilizes three types of cell-binding mechanisms: DOPA adhesion of Cell-Tak, cationic binding force of PLL, and RGD sequence-mediated adhesion of fibronectin. Therefore, the new recombinant fp-151-RGD is suitable for use as a cell-adhesion material in cell culture or tissue engineering, and in any other area where efficient cell adhesion is required.

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