Nano-liquid Chromatography-orbitrap MS-based Quantitative Proteomics Reveals Differences Between the Mechanisms of Action of Carnosic Acid and Carnosol in Colon Cancer Cells.
Carnosic acid (CA) and carnosol (CS) are two structurally related diterpenes present in rosemary herb (Rosmarinus officinalis). Although several studies have demonstrated that both diterpenes can scavenge free radicals and interfere in cellular processes such as cell proliferation, they may not necessarily exert the same effects at the molecular level. In this work, a shotgun proteomics study based on stable isotope dimethyl labeling (DML) and nano-liquid chromatography-tandem mass spectrometry (nano-LC-MS/MS) has been performed to identify the relative changes in proteins and to gain some light on the specific molecular targets and mechanisms of action of CA and CS in HT-29 colon cancer cells. Protein profiles revealed that CA and CS induce different Nrf2-mediated response. Furthermore, examination of our data revealed that each diterpene affects protein homeostasis by different mechanisms. CA treatment induces the expression of proteins involved in the unfolded protein response in a concentration dependent manner reflecting ER stress, whereas CS directly inhibits chymotrypsin-like activity of the 20S proteasome. In conclusion, the unbiased proteomics-wide method applied in the present study has demonstrated to be a powerful tool to reveal differences on the mechanisms of action of two related bioactive compounds in the same biological model.
From the ‡Laboratory of Foodomics, Institute of Food Science Research (CIAL, CSIC), Calle Nicolás Cabrera 9, 28049 Madrid, Spain.
Alberto
A
Valdés
From the ‡Laboratory of Foodomics, Institute of Food Science Research (CIAL, CSIC), Calle Nicolás Cabrera 9, 28049 Madrid, Spain; virginia.garcia@csic.es.
Virginia
0000-0001-6728-4325
V
García-Cañas
§Analytical Chemistry, Department of Chemistry-BMC and SciLifeLab, Uppsala University, Husargatan 3, 75124 Uppsala, Sweden.
Konstantin A
KA
Artemenko
From the ‡Laboratory of Foodomics, Institute of Food Science Research (CIAL, CSIC), Calle Nicolás Cabrera 9, 28049 Madrid, Spain.
Carolina
C
Simó
§Analytical Chemistry, Department of Chemistry-BMC and SciLifeLab, Uppsala University, Husargatan 3, 75124 Uppsala, Sweden.
Jonas
J
Bergquist
From the ‡Laboratory of Foodomics, Institute of Food Science Research (CIAL, CSIC), Calle Nicolás Cabrera 9, 28049 Madrid, Spain.
Alejandro
A
Cifuentes
D045784:Abietanes / Q000494:pharmacology*; D002853:Chromatography, Liquid / Q000379:methods*; D003110:Colonic Neoplasms / Q000378:metabolism*; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D019073:HT29 Cells; D006706:Homeostasis / Q000187:drug effects; D006801:Humans; D007553:Isotope Labeling; D046988:Proteasome Endopeptidase Complex / Q000187:drug effects; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction / Q000187:drug effects; D053719:Tandem Mass Spectrometry / Q000379:methods*; D056811:Unfolded Protein Response / Q000187:drug effects
D045784:Abietanes; C068623:carnosol; D046988:Proteasome Endopeptidase Complex; C018381:salvin