RESUMEN
The Clp/Hsp100 AAA+ chaperone family is involved in recovering aggregated proteins and little is known about other orthologs of the well studied ClpB from Escherichia coli and Hsp104 from Saccharomyces cerevisiae. Plant Hsp101 is a good model for understanding the relationship between the structure and function of Hsp100 proteins and to investigate the role of these chaperones in disaggregation processes. Here, we present the cloning and purification of a sugarcane ortholog, SHsp101, which is expressed in sugarcane cells and is a folded hexamer that is capable of binding nucleotides. Thus SHsp101 has the structural and functional characteristics of the Clp/Hsp100 AAA+ family.
Asunto(s)
Chaperonas Moleculares/metabolismo , Proteínas de Plantas/metabolismo , Proteínas Recombinantes/metabolismo , Saccharum/química , Factores de Transcripción/metabolismo , Secuencia de Aminoácidos , Quimera/genética , Clonación Molecular , Endopeptidasa Clp , Escherichia coli/química , Escherichia coli/genética , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Proteínas de Choque Térmico/química , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Espectroscopía de Resonancia Magnética , Modelos Moleculares , Chaperonas Moleculares/genética , Chaperonas Moleculares/aislamiento & purificación , Datos de Secuencia Molecular , Nucleótidos/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/aislamiento & purificación , Plásmidos , Polimerizacion , Unión Proteica , Pliegue de Proteína , Estructura Terciaria de Proteína , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharum/genética , Relación Estructura-Actividad , Factores de Transcripción/genética , Factores de Transcripción/aislamiento & purificación , UltracentrifugaciónRESUMEN
Several Gram-negative bacterial pathogens have developed type III secretion systems (T3SSs) to deliver virulence proteins directly into eukaryotic cells in a process essential for many diseases. The type III secretion processes require customized chaperones with high specificity for binding partners, thus providing the secretion to occur. Due to the very low sequence similarities among secretion chaperones, annotation and discrimination of a great majority of them is extremely difficult and a task with low scores even if genes are encountered that codify for small (<20 kDa) proteins with low pI and a tendency to dimerise. Concerning about this, herein, we present structural features on two hypothetical T3SSs chaperones belonging to plant pathogen Xanthomonas axonopodis pv. citri and suggest how low resolution models based on Small Angle X-ray Scattering patterns can provide new structural insights that could be very helpful in their analysis and posterior classification.
Asunto(s)
Proteínas Bacterianas/química , Chaperonas Moleculares/química , Xanthomonas axonopodis/química , Proteínas Bacterianas/metabolismo , Sistemas de Secreción Bacterianos , Dicroismo Circular , Escherichia coli , Modelos Moleculares , Chaperonas Moleculares/metabolismo , Conformación Proteica , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Dispersión del Ángulo Pequeño , Difracción de Rayos X , Xanthomonas axonopodis/metabolismoRESUMEN
Many Gram-negative bacteria are able to invade hosts by translocation of effectors directly into target cells in processes usually mediated by two very complex secretion systems (SSs), named type III (T3) and type IV (T4) SSs. These syringe-needle injection devices work with intervention of specialized secretion chaperones that, unlike traditional molecular chaperones, do not assist in protein folding and are not energized by ATP. Controversy still surrounds secretion chaperones primary role, but we can say that these chaperones act as: (i) bodyguards to prevent premature aggregation, or as (ii) pilots to direct substrate secretion through the correct secretion system. This family of chaperones does not share primary structure similarity but amazingly equal 3D folds. This mini review has the intent to present updated structural and functional data for several important secretion chaperones, either alone or in complex with their cognate substrates, as well to report on the common features and roles of T3, T4 and flagellar chaperones.
Asunto(s)
Sistemas de Secreción Bacterianos/fisiología , Bacterias Gramnegativas/fisiología , Chaperonas Moleculares/fisiología , Flagelos/fisiología , Chaperonas Moleculares/química , Conformación ProteicaRESUMEN
Metabolic profiling employing hydrogen nuclear magnetic resonance (1H NMR) spectroscopy and chemometric analysis of human blood serum samples taken from the control group (n = 25) and patients with bipolar disorder (n = 25) was performed to identify molecular changes related to the disorder and to different drug treatments: lithium (n = 15) versus other medications (n = 10). This strategy showed significant potential for exploring pathophysiological and toxicological features involved in bipolar disorder. The investigated groups (control and patients with bipolar disorder under different treatments) could be distinguished according to their metabolic profiles, and the main differential metabolites found were lipids, lipid-metabolism-related molecules (acetate, choline, and myo-inositol), and some key amino acids (glutamate, glutamine). Our results suggest that some of the 24 identified metabolites may be linked to lithium- and other-medication-provoked metabolic changes or may even be directly related to the disorder. Thus, these findings may contribute to paving the way for future studies aiming at identifying potential biomarkers for bipolar disorder.
