68 research outputs found
Estudio de variantes de la apolipoproteĂna A-I humana involucradas en amiloidosis
Las amiloidosis constituyen un grupo de enfermedades caracterizadas por la conversiĂłn de la estructura nativa de ciertas proteĂnas en una conformaciĂłn mal plegada que se dan generalmente con ganancia de estructura de hoja-beta, produciendo la agregaciĂłn y depĂłsito de estas proteĂnas lo que lleva al daño tisular, mal funcionamiento de los Ăłrganos, y en algunos casos conlleva a la muerte. El agregado de las proteĂnas se caracteriza por un marcado polimorfismo en donde oligĂłmeros, fibras, y agregados amorfos, se encuentran como producto final. Se ha reportado la presencia de Ia apoA-I humana (mayor constituyente proteico de las HDL) en lesiones del tipo amiloide, tanto la proteĂna wt como algunas mutantes puntuales. El presente trabajo propone caracterizar estructuralmente a las variantes de apoA-I Gly26Arg y Lys107-0 ambas asociadas con amiloidosis. Por otro lado se estudia como los microambientes celulares afectan las estabilidad de las proteĂnas en estudio favoreciendo el mal plegamiento que conduce a una agregaciĂłn patolĂłgica.Facultad de Ciencias Exacta
The trophoblast clock controls transport across placenta in mice
In mammals, 24-h rhythms of physiology and behavior are organized by a body-wide network of clock genes and proteins. Despite the well-known function of the adult circadian system, the roles of maternal, fetal and placental clocks during pregnancy are poorly defined. In the mature mouse placenta, the labyrinth zone (LZ) is of fetal origin and key for selective nutrient and waste exchange. Recently, clock gene expression has been detected in LZ and other fetal tissues; however, there is no evidence of a placental function controlled by the LZ clock. Here, we demonstrate that specifically the trophoblast layer of the LZ harbors an already functional clock by late gestation, able to regulate in a circadian manner the expression and activity of the xenobiotic efflux pump, ATP-binding cassette sub-family B member 1 (ABCB1), likely gating the fetal exposure to drugs from the maternal circulation to certain times of the day. As more than 300 endogenous and exogenous compounds are substrates of ABCB1, our results might have implications in choosing the maternal treatment time when aiming either maximal/minimal drug availability to the fetus/mother.Fil: Demarez, CĂ©cile. Universität Zu LĂĽbeck; AlemaniaFil: de Assis, Leonardo Vinicius Monteiro. Universität Zu LĂĽbeck; AlemaniaFil: Krohn, Markus. Universität Zu LĂĽbeck; AlemaniaFil: Ramella, Nahuel. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - La Plata. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias MĂ©dicas. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Schwaninger, Markus. Universität Zu LĂĽbeck; AlemaniaFil: Oster, Henrik. Universität Zu LĂĽbeck; AlemaniaFil: Astiz, Mariana. Universität Zu LĂĽbeck; Alemania. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas; Argentin
Dataset of the construction and characterization of stable biological nanoparticles
This article shows the dataset of clearance assays and the reconstitution of stable biological nano-complexes using both detergent-assisted and spontaneous solubilization of phospholipids by the recombinant purified apolipoprotein A-I (apoA-I). Protein was intra-chain crosslinked in order to introduce steric constrains. Then, native and crosslinked protein function was evaluated by a data collection of dimiristoyl phosphatidyl choline (DMPC) micellization curves. Additionally, resulting particles from spontaneous or detergent-assisted lipid solubilization were characterized by transmission electron microscopy (TEM), size exclusion chromatography (SEC), and native polyacrylamide gel electrophoresis (PAGE). Here we set up an experimental design that may help study protein structure based on its function, since interaction with biological membranes and lipids is an intrinsic activity attributed to many proteins in circulation. In addition, by t-test analysis of collected-data, we examined the formation of lipoprotein particles by native and intra-chain crosslinked proteins under different conditions like temperature and time incubation. Thus, data shown here strengthen the usefulness of an easy, rapid, accessible and inexpensive approach to test protein flexibility related to its function.Fil: Gisonno, Romina Antonela. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - La Plata. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias MĂ©dicas. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Tricerri, Maria Alejandra. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - La Plata. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias MĂ©dicas. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Gonzalez, Marina Cecilia. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - La Plata. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias MĂ©dicas. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Garda, Horacio Alberto. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - La Plata. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias MĂ©dicas. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Ramella, Nahuel. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - La Plata. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias MĂ©dicas. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: DĂaz Ludovico, Ivo. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - La Plata. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias MĂ©dicas. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentin
Aterosclerosis y amiloidosis: Âżdos patologĂas crĂłnicas interrelacionadas?
A fin de poder llevar a cabo las distintas funciones biológicas, es esencial que las proteínas conserven su conformación nativa. Algunas proteínas son estructuralmente inestables, y entonces pequeños cambios en el microambiente en el que se encuentran pueden ser clave para alterar el equilibrio hacia una conformación patológica. Las amiloidosis se caracterizan por la presencia de depósitos extracelulares de proteínas que adoptan estructura fibrilar. La apolipoproteína A-I humana no está normalmente asociada a esta patología, aunque fueron detectados agregados de la misma con la secuencia nativa en placas ateroscleróticas seniles. A pesar de ser frecuente, se conoce relativamente poco de la patogénesis y significancia de la agregación patológica de la apoA-I
Amiloidosis inducida por variantes de la apolipoproteĂna A-I humana
Si bien las amiloidosis severas no suelen ser patologĂas prevalentes, el plegamiento anĂłmalo de proteĂnas está involucrado a procesos crĂłnicos de inflamaciĂłn. Los niveles de apolipoprotein A-I (apoA-I) humana se correlacionan inversamente con el riesgo de sufrir enfermedades cardiovasculares por su participaciĂłn en el transporte reverso de colesterol; sin embargo, depĂłsitos de la forma nativa de la proteĂna se encuentran asociados a placas seniles, y algunas mutaciones puntuales inducen en paciente la formaciĂłn de depĂłsitos asociados a placas ateromatosas o disfunciĂłn renal.Facultad de Ciencias MĂ©dica
Data regarding the sensibility to proteolysis of a natural apolipoprotein A-I mutant
The article shows dataset of the proteolysis of a natural variant of apolipoprotein A-I (apoA-I) with a substitution of a leucine by and arginine in position 60 (L60R), in comparison with the protein with the native sequence (Wt). This information demonstrates the potential of in vitro partial proteolysis experiments as it may be applicable to different approaches in the biophysical field. We have analyzed by different electrophoresis techniques apoA-I variants, quantified the degree of proteolysis after staining and compared the proteolysis efficiency with the computed cleavage patterns. The data shown here clearly strengthen the usefulness of this approach to test protein flexibility, as it may be attained with enzymes which are not expected to modify in vivo this protein but have a well-known digestion pattern. In addition it is appropriate for evaluating protein catabolism, as it is exemplified here by the evidence with metalloproteinase 12 (MMP-12), which is a physiological protease that may elicit the pro-inflammatory processing of this variant within the lesions. We support the work “Structural analysis of a natural apolipoprotein A-I variant (L60R) associated with amyloidosis” (Gaddi, et al., 2020), gaining insights on protein folding from a characterization by proteolysis analysis [1].Fil: Gaddi, Gisela Marina. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - La Plata. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias MĂ©dicas. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Gisonno, Romina Antonela. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - La Plata. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias MĂ©dicas. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Rosu, Silvana Antonia. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - La Plata. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias MĂ©dicas. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Cortez, MarĂa Fernanda. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - La Plata. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias MĂ©dicas. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Finarelli, Gabriela Sandra. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - La Plata. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias MĂ©dicas. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Ramella, Nahuel. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - La Plata. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias MĂ©dicas. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Tricerri, Maria Alejandra. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - La Plata. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias MĂ©dicas. Instituto de Investigaciones BioquĂmicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentin
Amyloidogenic propensity of a natural variant of human apolipoprotein A-I: Stability and interaction with ligands
A number of naturally occurring mutations of human apolipoprotein A-I (apoA-I) have been associated with hereditary amyloidoses. The molecular mechanisms involved in amyloid-associated pathology remain largely unknown. Here we examined the effects of the Arg173Pro point mutation in apoA-I on the structure, stability, and aggregation propensity, as well as on the ability to bind to putative ligands. Our results indicate that the mutation induces a drastic loss of stability, and a lower efficiency to bind to phospholipid vesicles at physiological pH, which could determine the observed higher tendency to aggregate as pro-amyloidogenic complexes. Incubation under acidic conditions does not seem to induce significant desestabilization or aggregation tendency, neither does it contribute to the binding of the mutant to sodium dodecyl sulfate. While the binding to this detergent is higher for the mutant as compared to wt apoA-I, the interaction of the Arg173Pro variant with heparin depends on pH, being lower at pH 5.0 and higher than wt under physiological pH conditions. We suggest that binding to ligands as heparin or other glycosaminoglycans could be key events tuning the fine details of the interaction of apoA-I variants with the micro-environment, and probably eliciting the toxicity of these variants in hereditary amyloidoses.Facultad de Ciencias MĂ©dicasInstituto de Investigaciones BioquĂmicas de La PlataInstituto de Investigaciones FisicoquĂmicas TeĂłricas y Aplicada
Data related to inflammation and cholesterol deposition triggered by macrophages exposition to modified LDL
This article supports experimental evidence on the time-dependent effect on gene expression related to inflammation and cholesterol deposition in lipid-loaded cells. The cells employed were human monocytes THP1 line transformed into macrophages by treatment with phorbol esters. Macrophages were treated at different times with oxidized low density lipoprotein (Ox-LDL) and then gene expression was measured. We also include data about the different types of oxidized lipoprotein obtained (low, media or high oxidation) for differential exposure with Cu ions. These data include characterization to lipid and protein peroxidative damage and also quantification of cell viability by exposure to native and modified LDL. The present article complements data published in "Decreased OxLDL uptake and cholesterol efflux in THP1 cells elicited by cortisol and by cortisone through 11β-hydroxysteroid dehydrogenase type 1" Ledda et al. (in press) [. 1].Instituto de Investigaciones BioquĂmicas de La Plat
Human apolipoprotein A-I natural variants: molecular mechanisms underlying amyloidogenic propensity
Human apolipoprotein A-I (apoA-I)-derived amyloidosis can present with either wild-type (Wt) protein deposits in atherosclerotic plaques or as a hereditary form in which apoA-I variants deposit causing multiple organ failure. More than 15 single amino acid replacement amyloidogenic apoA-I variants have been described, but the molecular mechanisms involved in amyloid-associated pathology remain largely unknown. Here, we have investigated by fluorescence and biochemical approaches the stabilities and propensities to aggregate of two disease-associated apoA-I variants, apoA-IGly26Arg, associated with polyneuropathy and kidney dysfunction, and apoA-ILys107-0, implicated in amyloidosis in severe atherosclerosis. Results showed that both variants share common structural properties including decreased stability compared to Wt apoA-I and a more flexible structure that gives rise to formation of partially folded states. Interestingly, however, distinct features appear to determine their pathogenic mechanisms. ApoA-ILys107-0 has an increased propensity to aggregate at physiological pH and in a pro-inflammatory microenvironment than Wt apoA-I, whereas apoA-IGly26Arg elicited macrophage activation, thus stimulating local chronic inflammation. Our results strongly suggest that some natural mutations in apoA-I variants elicit protein tendency to aggregate, but in addition the specific interaction of different variants with macrophages may contribute to cellular stress and toxicity in hereditary amyloidosis
Especies intermedias en la agregaciĂłn de la apoliproteĂna A-I: probable rol en la amiloidosis
Las amiloidosis son patologĂas crĂłnicas, en las que proteĂnas mal plegadas inducen citotoxicidad de distinta severidad y en distintos Ăłrganos. Si bien gran cantidad de bibliografĂa sugiere que agregados fibrilares maduros serĂan la especie con mayor patogenicidad, estudios más recientes indican que oligĂłmeros o protofibras podrĂan jugar un rol fundamental en la cascada de eventos involucrados en la amiloidosis. La amiloidosis inducida por mutantes naturales de la apolipoproteĂna A-I humana (apoA-I) se presenta con diversas manifestaciones clĂnicas dependiendo de la variante proteica que se encuentra involucrada. En estudios previos determinamos que distintas variantes de esta proteĂna poseen aumentada tendencia a agregarse, y que parámetros pro-inflamatorios del micro ambiente favorecen un procesamiento patogĂ©nico. En este trabajo extendemos nuestros estudios, y nos planteamos resolver la hipĂłtesis de que distintas especies de agregaciĂłn intermedia participan en procesos citotĂłxicos mediados por variantes de apoA-I.Facultad de Ciencias MĂ©dica
- …