Person:
Bautista Santa Cruz, José Manuel

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First Name
José Manuel
Last Name
Bautista Santa Cruz
URI
https://hdl.handle.net/20.500.14352/80584
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Veterinaria
Department
Bioquímica y Biología Molecular
Area
Bioquímica y Biología Molecular
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Author: Azcárate, Isabel G. ×

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    Protein Susceptibility to Peroxidation by 4-Hydroxynonenal in Hereditary Hemochromatosis
    (International Journal of Molecular Sciences, 2023) Sánchez-Jaut, Sandra ; Pérez-Benavente, Susana ; Abad, Paloma ; Méndez Cuadro, Darío ; Puyet Catalina, Antonio; Díez Martín, Amalia; Galicia Poblet, Gonzalo ; Gómez Domínguez, Elena ; Moran Jiménez, María J. ; Bautista Santa Cruz, José Manuel; Azcárate, Isabel G.
    Iron overload caused by hereditary hemochromatosis (HH) increases free reactive oxygen species that, in turn, induce lipid peroxidation. Its 4-hydroxynonenal (HNE) by-product is a wellestablished marker of lipid peroxidation since it reacts with accessible proteins with deleterious consequences. Indeed, elevated levels of HNE are often detected in a wide variety of human diseases related to oxidative stress. Here, we evaluated HNE-modified proteins in the membrane of erythrocytes from HH patients and in organs of Hfe−/− male and female mice, a mouse model of HH. For this purpose, we used one- and two-dimensional gel electrophoresis, immunoblotting and MALDI-TOF/TOF analysis. We identified cytoskeletal membrane proteins and membrane receptors of erythrocytes bound to HNE exclusively in HH patients. Furthermore, kidney and brain of Hfe−/−mice contained more HNE-adducted protein than healthy controls. Our results identified main HNE-modified proteins suggesting that HH favours preferred protein targets for oxidation by HNE.
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    Shotgun Characterization of the Circulating IgM Antigenome of an Infectious Pathogen by Immunocapture-LC–MS/MS from Dried Serum Spots
    (Journal of Proteome Research, 2024) Abad González, Paloma; Coronado Brieva, Montserrat; Vincelle-Nieto, África; Pérez Benavente, Susana; Fobil, Julius N.; Puyet Catalina, Antonio; Díez Martín, Amalia; Reyes Palomares, Armando Adolfo; Azcárate, Isabel G.; Bautista Santa Cruz, José Manuel
    One of the main challenges in compiling the complete collection of protein antigens from pathogens for the selection of vaccine candidates or intervention targets is to acquire a broad enough representation of them to be recognized by the highly diversified immunoglobulin repertoire in human populations. Dried serum spot sampling (DSS) retains a large repertoire of circulating immunoglobulins from each individual that can be representative of a population, according to the sample size. In this work, shotgun proteomics of an infectious pathogen based on DSS sampling coupled with IgM immunoprecipitation, liquid chromatography–mass spectrometry (LC–MS/MS), and bioinformatic analyses was combined to characterize the circulating IgM antigenome. Serum samples from a malaria endemic region at different clinical statuses were studied to optimize IgM binding efficiency and antibody leaching by varying serum/immunomagnetic bead ratios and elution conditions. The method was validated using Plasmodium falciparum extracts identifying 110 of its IgM-reactive antigens while minimizing the presence of human proteins and antibodies. Furthermore, the IgM antigen recognition profile differentiated between malaria-infected and noninfected individuals at the time of sampling. We conclude that a shotgun proteomics approach offers advantages in providing a high-throughput, reliable, and clean way to identify IgM-recognized antigens from trace amounts of serum. The mass spectrometry raw data and metadata have been deposited with ProteomeXchange via MassIVE with the PXD identifier PXD043800.