Synthesis of biologically active tripeptide Arg- Gly-Asp (RGD).
Poster Number
7
Format
Poster Presentation
Abstract/Artist Statement
The short sequence like Arg-Gly-Asp (RGD) is primarily involved in protein binding on cell surfaces. Model studies using RGD and RGD-derived peptide are very important toward understanding the biological function of RGD-containing peptides. However RGD peptides are not easily available commercially. We synthesized the tripeptide RGD using solid-phase peptide synthesis (SPPS). The Wang Resin was used as the solid subport. The amino acids Asp, Gly and Arg were coupled to the Wang Resin, respectively. The coupling reagents used were diisoproplycarbodiimide (DIC) and 1- Hydroxybenzotriazole (HOBT). The amino acids used were protected with 9- fluorenylmethyloxycarbonyl (Fmoc) at the N-terminus. This N-terminus protecting group was removed by basic piperidine. An UV-Vis spectrophotometer was used to determine the progress of the deprotection reaction, for fmoc absorbs UV light at 301 nm. Once the entire peptide chain Arg-Gly-Asp was linked to the resin, the peptide was cleaved with Triflouroacetic acid (TFA). The solution obtained was concentrated with a rotovap. Chloroform extraction was performed to remove organic impurities. A crude peptide was obtained by removing the solvent using a centrifuge liked to a vacuum pump. The crude peptide was introduced in a mass spectrometer for MS/MS analysis.
Location
Pacific Geosciences Center
Start Date
24-4-2004 9:00 AM
End Date
24-4-2004 5:00 PM
Synthesis of biologically active tripeptide Arg- Gly-Asp (RGD).
Pacific Geosciences Center
The short sequence like Arg-Gly-Asp (RGD) is primarily involved in protein binding on cell surfaces. Model studies using RGD and RGD-derived peptide are very important toward understanding the biological function of RGD-containing peptides. However RGD peptides are not easily available commercially. We synthesized the tripeptide RGD using solid-phase peptide synthesis (SPPS). The Wang Resin was used as the solid subport. The amino acids Asp, Gly and Arg were coupled to the Wang Resin, respectively. The coupling reagents used were diisoproplycarbodiimide (DIC) and 1- Hydroxybenzotriazole (HOBT). The amino acids used were protected with 9- fluorenylmethyloxycarbonyl (Fmoc) at the N-terminus. This N-terminus protecting group was removed by basic piperidine. An UV-Vis spectrophotometer was used to determine the progress of the deprotection reaction, for fmoc absorbs UV light at 301 nm. Once the entire peptide chain Arg-Gly-Asp was linked to the resin, the peptide was cleaved with Triflouroacetic acid (TFA). The solution obtained was concentrated with a rotovap. Chloroform extraction was performed to remove organic impurities. A crude peptide was obtained by removing the solvent using a centrifuge liked to a vacuum pump. The crude peptide was introduced in a mass spectrometer for MS/MS analysis.