Proteins are effectors of most physiological functions in cells and they play a key role in the extracellular medium. Protein function is finely tuned at multiple levels including synthesis and degradation, intracellular localization and posttranslational modifications. The posttranslational modification of proteins is a key mechanism for the regulation of their biological activity. Various biological mediators, including lipids and reactive oxygen and nitrogen species, as well as some drugs and toxins, can modify proteins posttranslationally. We are interested in the study of the importance of these processes in molecular pathophysiology and in the mechanisms of drug action. Our work includes several aspects: the structural and functional characterization of novel types of posttranslational modifications, the identification of novel protein targets for modification and the study of the pathophysiological consequences of these phenomena.
Protein lipoxidation is the posttranslational modification of proteins by oxidized reactive lipids. This is a process with ample implications in physiology and pathophysiology, as illustrated in the figure below. We are exploring novel aspects of this modification.
Intermediate filament proteins of the type III family, including vimentin and GFAP, play key roles in cell dynamics, homeostasis and cytoskeletal cross-talk. We have identified these proteins as targets for modifications by electrophilic lipids and oxidants, preferentially at their single cysteine residue. This has led to define their role as redox and stress sensors. Moreover, we are delineating novel roles for these proteins in essential cellular processes, like the interaction of vimentin with actin in cell division, and elucidating their involvement in disease.
Modification of proteins by drugs or their metabolites, a process know as haptenation, can contribute to adverse reactions, including drug allergy. We are studying protein haptenation by several drugs in the context of a National network on "Asthma, adverse reactions and allergy".
Griesser E, Vemula V, Mónico A, Pérez-Sala D, Fedorova M. . Dynamic posttranslational modifications of cytoskeletal proteins unveil hot spots under nitroxidative stress. Redox Biol. 44, 102014.
Duarte S, Viedma-Poyatos A, Navarro-Carrasco E, Martínez, AE, Pajares, MA, Pérez-Sala D . Vimentin filaments interact with the actin cortex in mitosis allowing normal cell division. Nature Communications 10:4200
Mónico A, Guzmán-Caldentey J, Pajares MA, Martín-Santamaría S, Pérez-Sala D. . Molecular Insight into the Regulation of Vimentin by Cysteine Modifications and Zinc Binding. Antioxidants. 10:1039.
Monico A, Duarte S, Pajares MA, Pérez-Sala D . Vimentin disruption by lipoxidation and electrophiles: role of the cysteine residue and filament dynamics. Redox Biol. 23:101098. doi: 10.1016/j.redox.2019.101098
Ramos I, Stamatakis K, Oeste CL, Pérez-Sala D . Vimentin as a Multifaceted Player and Potential Therapeutic Target in Viral Infections. Int J Mol Sci 21:4675. doi: 10.3390/ijms21134675.
Viedma-Poyatos Á, Pajares MA, Pérez-Sala D . Type III intermediate filaments as targets and effectors of electrophiles and oxidants. Redox Biol 36:101582. doi: 10.1016/j.redox.2020.101582
Pajares MA, Zimmerman T, Sánchez-Gómez FJ, Ariza A, Torres MJ, Blanca M, Cañada FJ, Montañez MI, Pérez-Sala D . Amoxicillin Inactivation by Thiol-Catalyzed Cyclization Reduces Protein Haptenation and Antibacterial Potency. Front Pharmacol 11:189. doi: 10.3389/fphar.2020.00189.
Viedma-Poyatos A, de Pablo Y, Pekny M, Pérez-Sala D . The cysteine residue of glial fibrillary acidic protein is a critical target for lipoxidation and required for efficient network organization. Free Rad Biol Med. 20;120:380-394. DOI: 10.1016/j.freeradbiomed.2018.04.007
Sánchez-Gómez FJ, González-Morena J, Vida Y, Pérez-Inestrosa E, Blanca M, Torres MJ, Pérez-Sala D . Amoxicillin haptenates intracellular proteins that can be transported in exosomes to target cells. Allergy. 72:385-396. DOI: 10.1111/all.12958
Pajares MA, Pérez-Sala D . Mammalian sulfur amino acid metabolism: a nexus between redox regulation, nutrition, epigenetics and detoxification. Antioxid Redox Signal. 29:408-452. doi: 10.1089/ars.2017.7237
Pérez-Sala D, Oeste CL, Martínez AE, Carrasco, MJ, Garzón B, Cañada FJ . Vimentin filament organization and stress sensing depend on its single cysteine residue and zinc binding. Nat Commun 6:7287
Díez-Dacal B, Sánchez-Gómez FJ, Sánchez-Murcia PA, Milackova I, Zimmerman T, Ballekova J, García-Martín E, Agúndez JA, Gharbi S, Gago F, Stefek M, Pérez-Sala D . Molecular interactions and implications of aldose reductase inhibition by PGA1 and clinically used prostaglandins. Mol Pharmacol. 89:42-52
Pérez-Sala D, Domingues R . Lipoxidation targets: from basic mechanisms to pathophysiology. Redox Biol. 23:101208. Doi: 10.1016/j.redox.2019.101208
Oeste CL, Pinar M, Schink KO, Martínez-Turrión J, Stenmark H, Peñalva MA, Pérez-Sala D . An isoprenylation and palmitoylation motif promotes intraluminal vesicle delivery of proteins in cells from distant species. PLoS ONE 9(9): e107190
Oeste CL, Martínez-López M, Pérez-Sala D . Taking a lipidation-dependent path toward endolysosomes. Commun Integr Biol. 8:5, e1078041
"The astrocyte nanofilament system in Alexander disease – from molecules to function, uncovering new leads for therapy” EJPRD2019-256 “ALEXANDER”. European Joint Program on Rare Diseases
“Validation of vimentin as a co-receptor for SARS-CoV-2 and intervention strategies”. CSIC PTI Global Health (PIE 202020E223/CSIC-COVID-19-100)
"Proteoforms in pathophysiology and drug action: focus on intermediate filament proteins and selected drug targets" Ministerio de Ciencia e Innovación RTI-2018-097624-B-I00 (cofunded FEDER).
This project aims at the characterization of protein modification by electrophilic lipid mediators and drugs with a special focus on intermediate filaments.
"Red de Reacciones Adversas a Alergenos y Fármacos (RIRAAF). Instituto de Salud Carlos III, Ref. RD16/0006/0021 . Duración, desde: 01/01/2017 hasta: 31/12/2021 Financiación: 98.125,00 €. Investigadora responsable (CIB-Margarita Salas): Dra. Mª Dolores Pérez-Sala Gozalo. Este proyecto fue co-financiado por el Instituto de Salud Carlos III y fondos FEDER de la UE"
Our contribution to this Network is focused on the identification and characterization of protein targets for haptenation by drugs, mainly beta-lactam antibiotics, and in providing knowledge for the assessment of their implications in drug allergy.
COST Action CA19105 EpiLipidNET "Pan-European Network in Lipidomics and Epilipidomics"
Synthetic Cell Initiative https://www.syntheticcell.eu/
EU Project 675132 (H2020-MSCA-ITN-2015). Innovative Training Network. "MASS Spectrometry TRaining network for Protein Lipid adduct Analysis". MASSTRPLAN. Oct 2015-Sept 2019.
COST Action CM1001 "Chemistry of non-enzymatic protein modification: modulation of protein structure and function"
COST: Action TD1304 Zinc-Net: the Network for the Biology of Zinc
COST Action CA15214 EUROCELLNET "An integrative action for multidisciplinary studies on cellular structural networks"
- Red de Reacciones Adversas a Alergenos y Fármacos (RIRAAF) RETIC ISCIII. RD12/0013/0008
- "Protein tag for endo-lysosomal localization and degradation". D. Pérez-Sala, P. Boya, K, Stamatakis. P200802721. September 25, 2008.
- "Compounds with 2-cyclopentenone structure as inhibitors of AKR family enzymes". D. Pérez-Sala, B. Díez. P201030449. March 25, 2010.
- “Use of 5-carboxymethyl-3-mercapto-1,2,4-triazino-[5,6-b]indoles and their pharmaceutical composition” Milan Stefek, Ivana Milackova, Beatriz Díez Dacal, Dolores Pérez-Sala Gozalo, Marta Soltesova-Prnova. WO2015/057175 A, 23-4-2015.