The GAG molecules employed in this study are heparin, heparan sulfate, hyaluronic acid, and sulfated hyaluronic acid. Herein, we studied the influence of different glycosaminoglycan (i.e., glycan GAG) molecules on the conformation of a GDNF-derived peptide (GAG binding motif, sixteen amino acid residues at the N-terminus) using both experimental and theoretical studies. The activity of GDNF is highly dependent on the interaction with sulfated glycans which bind at the N-terminus consisting of 19 residues. Glial-cell-line-derived neurotrophic factor (GDNF) is a protein that has therapeutic potential in the treatment of Parkinson's disease and other neurodegenerative diseases. Taken together, our studies demonstrate that sCMC conjugated PG fibers can immobilize and retain function of cationic growth factors and hence show potential for use in various tissue engineering applications.
FOURIER SELF DECONVOLUTION ORIGIN PRO 8.5 FREE
Immobilized fibroblast growth factor-2 and connective tissue growth factor were bioactive and induced proliferation and fibrogenic differentiation of infrapatellar fat pad derived mesenchymal stem cells respectively with efficiency similar to or better than free growth factors. Transforming growth factor-β1 immobilized on the sCMC conjugated fibers was retained for at least 4 weeks with negligible release (3%). The fibrous scaffold bound cationic molecules, was cytocompatible and exhibited a remarkable morphological and functional stability. Inspired by the natural electrostatic interaction of cationic growth factors with anionic sulfated glycosaminoglycans in the extracellular matrix, we developed electrospun poly(hydroxybutyrate)/gelatin (PG) fibers conjugated with anionic sulfated carboxymethylcellulose (sCMC) to enable growth factor immobilization via electrostatic interaction for tissue engineering.
Sucrose octasulfate was also able to enhance the thermal stability of bFGF, but not to the same extent as heparin. Further, heparin was observed to greatly increase the thermal stability of bFGF, raising the T m by 25 ☌. The results indicate that the binding of heparin involves highly specific interactions. We have determined the effects of binding of various monomeric and polymeric, sulfated and nonsulfated glycosaminoglycans and carbohydrate compounds. This spectroscopically observable change has allowed us to probe the functional determinants necessary for heparin to bind to bFGF and to induce the observed conformational change. Further, sucrose octasulfate, a compound which mimics the effects of heparin biologically, was also observed to induce this same conformational change. Heparan sulfate, a component of the endothelial extracellular matrix, was also observed to bind to bFGF and induce a similar conformational change to that observed for heparin.
The observed spectral changes suggest that the conformational change is highly localized most likely in the β-turn regions of the bFGF molecule. Baseline must be subtracted before the analysis.The binding of heparin to basic fibroblast growth factor (bFGF) induces a small but highly reproducible conformational change observable in the amide I region of the protein's infrared spectrum. X data must be evenly spaced, otherwise the result may not make sense.Ģ. Click OK button to create the deconvolved result.ġ. Click Preview button to display the deconvolved spectrum.The larger the factor, the smoother the deconvolved spectrum, and the wider the deconvolved peaks. Smoothing factor must be between 0 and 1. Specify Smoothing Factor to smooth the deconvolved spectrum.Generally the larger Gamma is, the narrower the deconvolved peak becomes. Uncheck Auto checkbox to specify a custom value. The line shape will be used to deconvolve the spectrum. A default value is provided for Gamma, the line shape width.Click the Fourier Self-Deconvolution icon in the Apps Gallery window to open the dialog.An icon will appear in the Apps Gallery window. The position and integrated area for each peak are preserved in the deconvolved spectrum.ĭownload the file FourierSelfDeconvolution.opx, and then drag-and-drop onto the Origin workspace. This tool can be used to find overlapping peaks in the spectrum.
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