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Identification of Cyanobacterial Strains with Potential for the Treatment of Obesity-Related Co-Morbidities by Bioactivity, Toxicity Evaluation and Metabolite Profiling
Open AccessArticle

Application of Bioactive Thermal Proteome Profiling to Decipher the Mechanism of Action of the Lipid Lowering 132-Hydroxy-pheophytin Isolated from a Marine Cyanobacteria

1
Department of Clinical and Experimental Medicine, Cell Biology, Faculty of Medicine, Linköping University, 581 85 Linköping, Sweden
2
CIIMAR, Interdisciplinary Centre of Marine and Environmental Research, 4450-208 Matosinhos, Portugal
3
Department of Physiology, Faculty of Medicine and Nursing, University of the Basque Country UPV/EHU, 48940 Leioa, Spain
4
Department of Physiology, Ikerbasque, Faculty of Medicine and Nursing, University of the Basque Country UPV/EH, 48940 Leioa, Spain
*
Author to whom correspondence should be addressed.
Mar. Drugs 2019, 17(6), 371; https://doi.org/10.3390/md17060371
Received: 24 April 2019 / Revised: 7 June 2019 / Accepted: 15 June 2019 / Published: 21 June 2019
(This article belongs to the Special Issue Marine Natural Products and Obesity)
The acceleration of the process of understanding the pharmacological application of new marine bioactive compounds requires identifying the compound protein targets leading the molecular mechanisms in a living cell. The thermal proteome profiling (TPP) methodology does not fulfill the requirements for its application to any bioactive compound lacking chemical and functional characterization. Here, we present a modified method that we called bTPP for bioactive thermal proteome profiling that guarantees target specificity from a soluble subproteome. We showed that the precipitation of the microsomal fraction before the thermal shift assay is crucial to accurately calculate the melting points of the protein targets. As a probe of concept, the protein targets of 132-hydroxy-pheophytin, a compound previously isolated from a marine cyanobacteria for its lipid reducing activity, were analyzed on the hepatic cell line HepG2. Our improved method identified 9 protein targets out of 2500 proteins, including 3 targets (isocitrate dehydrogenase, aldehyde dehydrogenase, phosphoserine aminotransferase) that could be related to obesity and diabetes, as they are involved in the regulation of insulin sensitivity and energy metabolism. This study demonstrated that the bTPP method can accelerate the field of biodiscovery, revealing protein targets involved in mechanisms of action (MOA) connected with future applications of bioactive compounds. View Full-Text
Keywords: thermal proteome profiling; mechanisms of action; bioactive compound; label-free quantitative proteomics; marine biodiscovery thermal proteome profiling; mechanisms of action; bioactive compound; label-free quantitative proteomics; marine biodiscovery
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MDPI and ACS Style

Carrasco del Amor, A.; Freitas, S.; Urbatzka, R.; Fresnedo, O.; Cristobal, S. Application of Bioactive Thermal Proteome Profiling to Decipher the Mechanism of Action of the Lipid Lowering 132-Hydroxy-pheophytin Isolated from a Marine Cyanobacteria. Mar. Drugs 2019, 17, 371. https://doi.org/10.3390/md17060371

AMA Style

Carrasco del Amor A, Freitas S, Urbatzka R, Fresnedo O, Cristobal S. Application of Bioactive Thermal Proteome Profiling to Decipher the Mechanism of Action of the Lipid Lowering 132-Hydroxy-pheophytin Isolated from a Marine Cyanobacteria. Marine Drugs. 2019; 17(6):371. https://doi.org/10.3390/md17060371

Chicago/Turabian Style

Carrasco del Amor, Ana; Freitas, Sara; Urbatzka, Ralph; Fresnedo, Olatz; Cristobal, Susana. 2019. "Application of Bioactive Thermal Proteome Profiling to Decipher the Mechanism of Action of the Lipid Lowering 132-Hydroxy-pheophytin Isolated from a Marine Cyanobacteria" Mar. Drugs 17, no. 6: 371. https://doi.org/10.3390/md17060371

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