Efficient Screening of Marine Extracts for Protease Inhibitors by Combining FRET Based Activity Assays and Surface Plasmon Resonance Spectroscopy Based Binding Assays
Abstract
:1. Introduction
2. Results and Discussion
2.1. Screening for Inhibitors of HIV-1 Protease, SAP1, SAP2, SAP3 and Pepsin
% Inhibition | |||||||
---|---|---|---|---|---|---|---|
Extract | HIV-1 protease | SAP1 | SAP2 | SAP3 | Pepsin | BACE1 | HCMV Protease |
P1-10 | 27 ± 1 | 11 ± 1 | −5 ± 6 | −6 ± 1 | 5 ± 2 | 7 ± 1 | 41 ± 4 |
P1-20 | 70 ± 13 | 47 ± 1 | 36 ± 15 | 44 ± 1 | 34 ± 2 | 44 ± 3 | 71 ± 4 |
P1-50 | 56 ± 6 | 75 ± 11 | 68 ± 4 | 76 ± 3 | 47 ± 13 | 27 ± 3 | 68 ± 10 |
P1-80 | −1 ± 1 | 29 ± 4 | 60 ± 1 | 51 ± 1 | 54 ± 14 | 2 ± 4 | 45 ± 5 |
P2-4 | 11 ± 1 | 10 ± 7 | 4 ± 11 | 6 ± 4 | 11 ± 11 | 3 ± 3 | 43 ± 6 |
P2-10 | 14 ± 3 | 21 ± 8 | −5 ± 4 | 8 ± 4 | 10 ± 5 | 11 ± 3 | 49 ± 2 |
P2-20 | 28 ± 3 | −5 ± 15 | 7 ± 1 | −2 ± 7 | 12 ± 1 | 22 ± 4 | 30 ± 9 |
P2-50 | −18 ± 4 | 8 ± 5 | 36 ± 13 | 14 ± 1 | 13 ± 6 | 9 ± 1 | 10 ± 3 |
2.2. Screening for Inhibitors of BACE1
2.3. Screening for Inhibition of HCMV Protease
3. Experimental Section
3.1. Preparation of Extracts from Norwegian Spring Spawning Herring
3.2. Protease Production and FRET Based Activity Assay
3.2.1. HIV-1 Protease
3.2.2. SAP1, SAP2 and SAP3
3.2.3. Pepsin
3.2.4. BACE1
3.2.5. HCMV Protease
3.3. SPR Based Binding Assays
3.3.1. HIV-1 Protease
3.3.2. SAP1, SAP2 and SAP3
3.3.3. BACE1
3.3.4. HCMV Protease
4. Conclusions
Acknowledgments
Conflicts of Interest
References
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Christopeit, T.; Øverbø, K.; Danielson, U.H.; Nilsen, I.W. Efficient Screening of Marine Extracts for Protease Inhibitors by Combining FRET Based Activity Assays and Surface Plasmon Resonance Spectroscopy Based Binding Assays. Mar. Drugs 2013, 11, 4279-4293. https://doi.org/10.3390/md11114279
Christopeit T, Øverbø K, Danielson UH, Nilsen IW. Efficient Screening of Marine Extracts for Protease Inhibitors by Combining FRET Based Activity Assays and Surface Plasmon Resonance Spectroscopy Based Binding Assays. Marine Drugs. 2013; 11(11):4279-4293. https://doi.org/10.3390/md11114279
Chicago/Turabian StyleChristopeit, Tony, Kersti Øverbø, U. Helena Danielson, and Inge W. Nilsen. 2013. "Efficient Screening of Marine Extracts for Protease Inhibitors by Combining FRET Based Activity Assays and Surface Plasmon Resonance Spectroscopy Based Binding Assays" Marine Drugs 11, no. 11: 4279-4293. https://doi.org/10.3390/md11114279