Project Title

Chromatographic Method for Antiviral Peptide Temporin L Interactions with the Main Protease of SARS-CoV-2

Academic department under which the project should be listed

CSM - Chemistry and Biochemistry

Faculty Sponsor Name

Mohammad A. Halim

Abstract (300 words maximum)

Temporin L (TL) is an antimicrobial peptide isolated from the skin of the European red frog Rana temporaria. Frog skin is a potential source of antimicrobial peptides which offer innate immunity and protection from various pathogenic microorganism. One of our recent studies showed that TL has antiviral properties with an inhibition efficiency against the main protease of SARS-CoV-2. Various techniques can be used to detect the protein-peptide interaction including classic protease assay, gel filtrations and affinity chromatography. In this study, we monitor the peptide-protein interactions without immobilized or chemical derivatization. All experiments were performed using an Agilent 1290 infinity II UHPLC system equipped with a binary pump, a multisampler and a UV detector excitation at 280 nm. ZORBAX C18 column was used for this study. The peptide-protein samples were equilibrated for 30 mins before injecting in the UPLC system. The concentration of the main protease was fixed at 10 µM while TL peptide concentration was varied from 5 µM to 50 µM. When protein sample was injected, it showed a single peak with an area of 228 mAU·S. The protein peak area is significantly reduced to 113 mAU·S when the protein sample with 5 µM TL was injected. In addition, a small shoulder peak is observed although the protein main peak was predominant. The shoulder peak size is significantly increased when protein sample with 10µM and 25 µM TL were injected. In high concentration of TL (50µM), the shoulder peak became the predominant while the protein peak is diminished remarkably. These results showed that the peptide binding induced conformational change of the main protease is evident from the preferential detection of the prominent shoulder peak when TL concentration is increased.

Disciplines

Analytical Chemistry | Biochemistry | Medicinal-Pharmaceutical Chemistry

Project Type

Poster

How will this be presented?

Yes, in person

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Chromatographic Method for Antiviral Peptide Temporin L Interactions with the Main Protease of SARS-CoV-2

Temporin L (TL) is an antimicrobial peptide isolated from the skin of the European red frog Rana temporaria. Frog skin is a potential source of antimicrobial peptides which offer innate immunity and protection from various pathogenic microorganism. One of our recent studies showed that TL has antiviral properties with an inhibition efficiency against the main protease of SARS-CoV-2. Various techniques can be used to detect the protein-peptide interaction including classic protease assay, gel filtrations and affinity chromatography. In this study, we monitor the peptide-protein interactions without immobilized or chemical derivatization. All experiments were performed using an Agilent 1290 infinity II UHPLC system equipped with a binary pump, a multisampler and a UV detector excitation at 280 nm. ZORBAX C18 column was used for this study. The peptide-protein samples were equilibrated for 30 mins before injecting in the UPLC system. The concentration of the main protease was fixed at 10 µM while TL peptide concentration was varied from 5 µM to 50 µM. When protein sample was injected, it showed a single peak with an area of 228 mAU·S. The protein peak area is significantly reduced to 113 mAU·S when the protein sample with 5 µM TL was injected. In addition, a small shoulder peak is observed although the protein main peak was predominant. The shoulder peak size is significantly increased when protein sample with 10µM and 25 µM TL were injected. In high concentration of TL (50µM), the shoulder peak became the predominant while the protein peak is diminished remarkably. These results showed that the peptide binding induced conformational change of the main protease is evident from the preferential detection of the prominent shoulder peak when TL concentration is increased.

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