Marta Vilaseca - Academia.edu (original) (raw)

Papers by Marta Vilaseca

British Journal of Pharmacology, Oct 1, 2000

The eects of supernatant from the bacterial strain Serratia marcescens 2170 (CS-2170) on the viab... more The eects of supernatant from the bacterial strain Serratia marcescens 2170 (CS-2170) on the viability of dierent haematopoietic cancer cell lines (Jurkat, NSO, HL-60 and Ramos) and nonmalignant cells (NIH-3T3 and MDCK) was studied. We examined whether this cytotoxic eect was due to apoptosis, and we puri®ed the molecule responsible for this eect and determined its chemical structure. 2 Using an MTT assay we showed a rapid (4 h) decrease in the number of viable cells. This cytotoxic eect was due to apoptosis, according to the fragmentation pattern of DNA, Hoechst 33342 staining and FACS analysis of the phosphatidylserine externalization. This apoptosis was blocked by using the caspase inhibitor Z-VAD.fmk, indicating the involvement of caspases. 3 Prodigiosin is a red pigment produced by various bacteria including S. marcescens. Using mutants of S. marcescens (OF, WF and 933) that do not synthesize prodigiosin, we further showed that prodigiosin is involved in this apoptosis. This evidence was corroborated by spectroscopic analysis of prodigiosin isolated from S. marcescens. 4 These results indicate that prodigiosin, an immunosuppressor, induces apoptosis in haematopoietic cancer cells with no marked toxicity in nonmalignant cells, raising the possibility of its therapeutic use as an antineoplastic drug.

Proteómica: revista de la Sociedad Española de Proteómica, 2013

Journal of Proteomics, 2017

Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic proteomic search engine ... more Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic proteomic search engine for the separation, detection, and identification of N-and O-glycopeptide glycoforms in standard glycoproteins. The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. Moreover, we evaluated several acquisition strategies in order to improve the detection and characterization of glycopeptide glycoforms with the maximum number of identification percentages. The proposed strategy is simple to set up with the technology platforms commonly used in proteomic labs. The method allows the straightforward and rapid obtention of a general glycosylated map of a given protein, including glycosites and their corresponding glycosylated structures. The MS strategy selected in this work, based on a gas phase fractionation approach, led to 136 unique peptides from four standard proteins, which represented 78% of the total number of peptides identified. Moreover, the method does not require an extra glycopeptide enrichment step, thus preventing the bias that this step could cause towards certain glycopeptide species. Data are available via ProteomeXchange with identifier PXD003578 Highlights Baseline-resolution of glycopeptide glycoforms on the basis of sialic acids Automatic and rapid identification of protein glycosites and glycan structures MS strategy for glycopeptide enrichment without additional experimental steps

Talanta, Nov 1, 2015

Transthyretin (TTR) is a homotetrameric protein which is known to misfold and aggregate causing d... more Transthyretin (TTR) is a homotetrameric protein which is known to misfold and aggregate causing different types of amyloidosis, such as familial amyloidotic polyneuropathy type I (FAP-I). FAP-I is associated with a specific TTR mutant variant (TTR (Met30)) that can be easily detected analysing the monomeric forms of the mutant protein. Meanwhile, the mechanism of protein aggregation onset, which could be triggered by structural changes on the native tetrameric protein complex, remains uncertain. We developed and described herein a new sample pretreatment based on immunoprecipitation (IP) to purify TTR from serum under non-denaturing conditions. Later, a nano-electrospray ionization-ion mobility mass spectrometry (nano-ESI-IM-MS or IM-MS) method was optimised to analyse the protein complexes in serum samples from healthy controls and FAP-I patients. IM-MS allowed separation and characterisation of tetrameric, trimeric and dimeric TTR gas ions due to their differential drift time, which is related to ion size and charge. The tetramer-to-dimer abundance ratio was differential between healthy controls and FAP-I patients (asymptomatic, symptomatic and an iatrogenic patient originally without the mutation who received a liver transplant from an FAP-I patient), and was also indicative of the effectiveness of liver transplantation as a treatment for FAP-I.

Journal of Peptide Science, Mar 1, 1999

DKP formation is a serious side reaction during the solid-phase synthesis of peptide acids contai... more DKP formation is a serious side reaction during the solid-phase synthesis of peptide acids containing either Pro or Gly at the C-terminus. This side reaction not only leads to a lower overall yield, but also to the presence in the reaction crude of several deletion peptides lacking the first amino acids. For the preparation of protected peptides using the Fmoc/tBu strategy, the use of a ClTrt-Cl-resin with a limited incorporation of the C-terminal amino acid is the method of choice. The use of resins with higher loading levels leads to more impure peptide crudes. The use of HPLC-ESMS is a useful method for analysing complex samples, such as those formed when C-terminal Pro peptides are prepared by non-optimized solid-phase strategies.

Talanta, May 1, 2018

In this study, we describe a chemometric data analysis approach to assist in the interpretation o... more In this study, we describe a chemometric data analysis approach to assist in the interpretation of the complex datasets from the analysis of high-molecular mass oligomeric proteins by ion mobility mass spectrometry (IM-MS). The homotetrameric protein transthyretin (TTR) is involved in familial amyloidotic polyneuropathy type I (FAP-I). FAP-I is associated with a specific TTR mutant variant (TTR(Met30)) that can be easily detected analyzing the monomeric forms of the mutant protein. However, the principal component analysis; PLS-DA, partial least squares discriminant analysis; TTR, transthyretin; VIP, variable importance in the projection.

European Journal of Organic Chemistry, Aug 1, 2002

ABSTRACT The synthesis of a set of [(azolio)methyl]azolate betaines 3−6 designed by combination o... more ABSTRACT The synthesis of a set of [(azolio)methyl]azolate betaines 3−6 designed by combination of a variety of heterocyclic frag-ments based on pyrazole, 1,2,4-triazole, and benzimidazole is reported. The dipolar nature of the betaines is discussed on the basis of 1 H and 13 C NMR spectroscopic data and dipole moment values, which range between 13.4 and 16.5 D. By exploitation of the sensitivity of electrospray ionization mass spectrometry in both the positive and negative modes,

Journal of Molecular Biology, May 1, 2019

The INhibitor of Growth (ING) family of tumor suppressors regulates the transcriptional state of ... more The INhibitor of Growth (ING) family of tumor suppressors regulates the transcriptional state of chromatin by recruiting remodeling complexes to sites with histone H3 trimethylated at lysine 4 (H3K4me3). This modification is recognized by the plant homeodomain (PHD) present at the C-terminus of the five ING proteins. ING5 facilitates histone H3 acetylation by the HBO1 complex, and also H4 acetylation by the MOZ/ MORF complex. We show that ING5 forms homodimers through its N-terminal domain, which folds independently into an elongated coiled-coil structure. The central region of ING5, which contains the nuclear localization sequence, is flexible and disordered, but it binds dsDNA with micromolar affinity. NMR analysis of the full-length protein reveals that the two PHD fingers of the dimer are chemically equivalent and independent of the rest of the molecule, and they bind H3K4me3 in the same way as the isolated PHD. We have observed that ING5 can form heterodimers with the highly homologous ING4, and that two of three primary tumorassociated mutants in the N-terminal domain strongly destabilize the coiled-coil structure. They also affect cell proliferation and cell cycle phase distribution, suggesting a driver role in cancer progression.

Rapid Communications in Mass Spectrometry, 2000

Electrospray ionization mass spectral analysis of simple dicationic imidazolium prototypes (M.2X)... more Electrospray ionization mass spectral analysis of simple dicationic imidazolium prototypes (M.2X) is reported and direct observation was obtained for proton-mediated ion-molecule reactions in the gas phase. The comparative ESI-MS study with heterophanes 1a.2Cl and 3.2Br, the open chain system 5.2Br, and their regiospecific deuterated counterparts 2a.2Cl, 4.2Br and 6.2Br showed that the nature of the frameworks containing two imidazolium subunits modulates their electrospray ionization, changing the stability of the common characteristic peaks: the carbene specie [M - H](+) or [M - D](+) as well as [M](2+) and [M. X](+).Copyright 2000 John Wiley & Sons, Ltd.

Tetrahedron, Dec 1, 1998

The reduction of methionine sulfoxide with ammonium iodide in trifluoroacetic acid has been studi... more The reduction of methionine sulfoxide with ammonium iodide in trifluoroacetic acid has been studied in peptides containing cysteine, histidine, tyrosine or tryptophan residues. While histidine and tyrosine have proved to be stable under the experimental conditions, cysteine is oxidized to cystine and tryptophan dimerizes to form 2-indolylindolenine derivatives. The use of methyl sulfide to increase the reduction rate minimizes the

Analytical Chemistry, Mar 14, 2018

Brain-derived amyloid-β (Aβ) dimers are associated with Alzheimer´s disease (AD). However, their ... more Brain-derived amyloid-β (Aβ) dimers are associated with Alzheimer´s disease (AD). However, their covalent nature remains controversial. This feature is relevant, as a covalent cross-link would make brain-derived dimers (brain dimers) more synaptotoxic than Aβ monomers and would make them suitable candidates for biomarker development. To resolve this controversy, we here present a three-step approach. First, we validated a type of synthetic cross-linked Aβ (CL Aβ) dimers, obtained by means of the photo-induced cross-linking of unmodified proteins (PICUP) reaction, as well-defined mimics of putative brain CL Aβ dimers. Second, we used these PICUP CL Aβ dimers as standards to improve the isolation of brain Aβ dimers and to develop state-of-the-art mass spectrometry (MS) strategies to allow their characterization. Third, we applied these MS methods to the analysis of brain Aβ dimer samples allowing the detection of the CL [Aβ(6-16)] 2 peptide comprising a dityrosine cross-link. This result demonstrates the presence of CL Aβ dimers in the brains of patients with AD and opens up avenues for establishing new therapeutic targets and developing novel biomarkers for this disease.

$Research paper thumbnail of Imidazolium-Based Dicationic Cyclophanes. Solid-State Aggregates with Unconventional (C–H)<sup>+</sup>···Cl<sup>−</sup> Hydrogen Bonding Revealed by X-ray Diffraction$

Supramolecular Chemistry, Oct 1, 2007

The first single-crystal X-ray crystallographic diffraction analysis of a dicationic heterophane ... more The first single-crystal X-ray crystallographic diffraction analysis of a dicationic heterophane showed a non-classic (C−H) + •••Cl − hydrogen bond between the imidazolium rings and halide anions and the formation of unconventional charged assisted hydrogen bonds, which were the non-covalent forces driving the anion interactions shown by the dications 4•2X. Here we report the halide-templated controlled synthesis and chemical response in basic media of 4•2X. Their structural properties were examined at the gas phase by electrospray ionization mass spectrometry in the negative-ion mode and in the solid-state by X-ray crystallography. Thus, the negative-ion ESI-MS response showed that the formation of non-covalent self-aggregates of macrocyclic dications is a consequence of hydrogen-bonded complexes with halide anions. Notably, X-ray diffraction of dication 4a•2Cl•2H 2 O provides evidence for the H-bonding network, which has a crucial role in crystal

Tetrahedron, May 1, 1995

Methionine sulfoxide reduction by NH4I has been studied in some disulfide containing peptides. In... more Methionine sulfoxide reduction by NH4I has been studied in some disulfide containing peptides. In general, this reagent has proved to be effective in neat TFA at 0°C, with the obtention of the unprotected peptides in more than 99% yields and without reduction of the disulfide bridge bond. The use of Me2S as an additive resulted in faster reactions and disulfide scrambling was not observed. Whereas the Acm group proved to be stable to the reaction conditions, unprotected cysteine containing peptides afforded the corresponding parallel dimers.

Science Advances, Aug 2, 2019

The notable male predominance across many human cancer types remains unexplained. Here, we show t... more The notable male predominance across many human cancer types remains unexplained. Here, we show that Drosophila l(3)mbt brain tumors are more invasive and develop as malignant neoplasms more often in males than in females. By quantitative proteomics, we have identified a signature of proteins that are differentially expressed between male and female tumor samples. Prominent among them is the conserved chromatin reader PHD finger protein 7 (Phf7). We show that Phf7 depletion reduces sex-dependent differences in gene expression and suppresses the enhanced malignant traits of male tumors. Our results identify potential regulators of sex-linked tumor dimorphism and show that these genes may serve as targets to suppress sex-linked malignant traits.

Investigational New Drugs, Apr 5, 2006

Prodigiosin (PG) is a bacterial, red-pigmented antibiotic with immunosuppressive and apoptotic ac... more Prodigiosin (PG) is a bacterial, red-pigmented antibiotic with immunosuppressive and apoptotic activities. To better understand its mechanisms of action, we tried to identify proteins associated with apoptosis induced by PG. For this purpose, the variation of protein expression on exposure to apoptotic concentrations of PG was examined, by high-resolution two-dimensional gel electrophoresis (2D-E), in the MCF-7 cancer cell line resistant to mitoxantrone (MCF-7-MR). Six PG apoptosis-associated protein spots were further characterized by complementary peptide mass fingerprinting and tandem mass spectrometry data obtained on a matrix-assisted laser desorption ionization-time-of-flight/time-of-flight (MALDI-TOF/TOF) mass spectrometer. The proteins identified were involved in various cellular functions, including cell defence, DNA repair and cellular organization. Our data provide novel information on cell response to PG, a new apoptotic drug with interesting anticancer activity.

Nature Methods, Aug 26, 2019

Respiratory Research

Background Premature birth, perinatal inflammation, and life-saving therapies such as postnatal o... more Background Premature birth, perinatal inflammation, and life-saving therapies such as postnatal oxygen and mechanical ventilation are strongly associated with the development of bronchopulmonary dysplasia (BPD); these risk factors, alone or combined, cause lung inflammation and alter programmed molecular patterns of normal lung development. The current knowledge on the molecular regulation of lung development mainly derives from mechanistic studies conducted in newborn rodents exposed to postnatal hyperoxia, which have been proven useful but have some limitations. Methods Here, we used the rabbit model of BPD as a cost-effective alternative model that mirrors human lung development and, in addition, enables investigating the impact of premature birth per se on the pathophysiology of BPD without further perinatal insults (e.g., hyperoxia, LPS-induced inflammation). First, we characterized the rabbit’s normal lung development along the distinct stages (i.e., pseudoglandular, canalicul...

An entry from the Cambridge Structural Database, the world's repository for small molecule cr... more An entry from the Cambridge Structural Database, the world's repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.