YocM a small heat shock protein can protect Bacillus subtilis cells during salt stress

Small heat shock proteins (sHsp) occur in all domains of life. By interacting with misfolded or aggregated proteins these chaperones fulfill a protective role in cellular protein homeostasis. Here, we demonstrate that the sHsp YocM of the Gram‐positive model organism Bacillus subtilis is part of the cellular protein quality control system with a specific role in salt stress response. In the absence of YocM the survival of salt shocked cells is impaired, and increased levels of YocM protect B. subtilis exposed to heat or salt. We observed a salt and heat stress‐induced localization of YocM to intracellular protein aggregates. Interestingly, purified YocM appears to accelerate protein aggregation of different model substrates in vitro. In addition, the combined presence of YocM and chemical chaperones, which accumulate in salt stressed cells, can facilitate in vitro a synergistic protective effect on protein misfolding. Therefore, the beneficial role of YocM during salt stress could be related to a mutual functional relationship with chemical chaperones and adds a new possible functional aspect to sHsp chaperone activities.     

Potential Relevance of α1-Adrenergic Receptor Autoantibodies in Refractory Hypertension

Background

Agonistic autoantibodies directed at the α₁-adrenergic receptor (α₁-AAB) have been described in patients with hypertension. We implied earlier that α₁-AAB might have a mechanistic role and could represent a therapeutic target.

Methodology/Principal Findings

To pursue the issue, we performed clinical and basic studies. We observed that 41 of 81 patients with refractory hypertension had α₁-AAB; after immunoadsorption blood pressure was significantly reduced in these patients. Rabbits were immunized to generate α₁-adrenergic receptor antibodies (α₁-AB). Patient α₁-AAB and rabbit α₁-AB were purified using affinity chromatography and characterized both by epitope mapping and surface plasmon resonance measurements. Neonatal rat cardiomyocytes, rat vascular smooth muscle cells (VSMC), and Chinese hamster ovary cells transfected with the human α_1A-adrenergic receptor were incubated with patient α₁-AAB and rabbit α₁-AB and the activation of signal transduction pathways was investigated by Western blot, confocal laser scanning microscopy, and gene expression. We found that phospholipase A2 group IIA (PLA2-IIA) and L-type calcium channel (Cacna1c) genes were upregulated in cardiomyocytes and VSMC after stimulation with both purified antibodies. We showed that patient α₁-AAB and rabbit α₁-AB result in protein kinase C alpha activation and transient extracellular-related kinase (EKR1/2) phosphorylation. Finally, we showed that the antibodies exert acute effects on intracellular Ca²⁺ in cardiomyocytes and induce mesentery artery segment contraction.

Conclusions/Significance

Patient α₁-AAB and rabbit α₁-AB can induce signaling pathways important for hypertension and cardiac remodeling. Our data provide evidence for a potential clinical relevance for α₁-AAB in hypertensive patients, and the notion of immunity as a possible cause of hypertension.     

A pathogenic PrP mutation and doppel interfere with polarized sorting of the prion protein

Several proteins linked to neurodegenerative diseases, such as the beta-amyloid precursor protein, amyloid beta-peptide, beta-secretase, and tau, undergo selective polarized sorting. We investigated polarized sorting of the mammalian prion protein (PrP(C)) and its homologue doppel (Dpl). In contrast to Dpl, which accumulates on the apical surface, PrP(C) is targeted selectively to the basolateral side in Madin-Darby canine kidney cells. An extensive deletion and domain swapping analysis revealed that the internal hydrophobic domain (HD) of PrP (amino acids 113-133) confers basolateral sorting in a dominant manner. PrP mutants lacking the HD are sorted apically, while Dpl chimeras containing the HD of PrP are directed to the basolateral membrane. Furthermore, a pathogenic PrP missense mutation within the HD leads to aberrant apical sorting of PrP as well.     

Excretion of metabolites by hydrogen bacteria

Summary D(–)-3-hydroxybutanoic acid was produced by a double mutant of Alcaligenes eutrophus, unable to synthesize poly-3-hydroxybutanoic acid and to utilize 3-hydroxybutanoate as a substrate. About 3.4 g of 3-hydroxybutanoate/l were produced under optimum conditions at pH 7.0 to 7.3, at a temperature of 30°C after exhaustion of ammonia and at restricted aeration rates allowing 10–12% of the maximum respiratory rate of the cells to occur. D(–)-3-hydroxybutanoic acid was identified by means of gas and ion exchange chromatography, IR-spectrometry and specific rotation.