Wrapping the alpha-crystallin domain fold in a chaperone assembly

Small heat shock proteins (sHsps) are oligomers that perform a protective function by binding denatured proteins. Although ubiquitous, they are of variable sequence except for a C-terminal similar to 90-residue "alpha-crystallin domain". Unlike larger stress response chaperones, sHsps are ATP-independent and generally form polydisperse assemblies. One proposed mechanism of action involves these assemblies breaking into smaller subunits in response to stress, before binding unfolding substrate and reforming into larger complexes. Two previously solved non-metazoan sHsp multimers are built from dimers formed by domain swapping between the alpha-crystallin domains,. adding to evidence that the smaller subunits are dimers. Here, the 2.5 angstrom resolution structure of an sHsp from the parasitic flatworm Taenia saginata Tsp36, the first metazoan crystal structure, shows a new mode of dimerization involving N-terminal regions, which differs from that seen for non-metazoan sHsps. Sequence differences in the a-crystallin domains between metazoans and nonmetazoans are critical to the different mechanism of dimerization, suggesting that some structural features seen for Tsp36 may be generalized to other metazoan sHsps. The structure also indicates scope for flexible assembly of subunits, supporting the proposed process of oligomer breakdown, substrate binding and reassembly as the chaperone mechanism. It further shows how sHsps can bind coil and secondary structural elements by wrapping them around the alpha-crystallin domain. The structure also illustrates possible roles for conserved residues associated with disease, and suggests a mechanism for the sHsp-related pathogenicity of some flatworm infections. Tsp36, like other flatworm sHsps, possesses two divergent sHsp repeats per monomer. Together with the two previously solved structures, a total of four alpha-crystallin domain structures are now available, giving a better definition of domain boundaries for sHsps

A redox switch in angiotensinogen modulates angiotensin release.

Blood pressure is critically controlled by angiotensins, which are vasopressor peptides specifically released by the enzyme renin from the tail of angiotensinogen-a non-inhibitory member of the serpin family of protease inhibitors. Although angiotensinogen has long been regarded as a passive substrate, the crystal structures solved here to 2.1 Å resolution show that the angiotensin cleavage site is inaccessibly buried in its amino-terminal tail. The conformational rearrangement that makes this site accessible for proteolysis is revealed in our 4.4 Å structure of the complex of human angiotensinogen with renin. The co-ordinated changes involved are seen to be critically linked by a conserved but labile disulphide bridge. Here we show that the reduced unbridged form of angiotensinogen is present in the circulation in a near 40:60 ratio with the oxidized sulphydryl-bridged form, which preferentially interacts with receptor-bound renin. We propose that this redox-responsive transition of angiotensinogen to a form that will more effectively release angiotensin at a cellular level contributes to the modulation of blood pressure. Specifically, we demonstrate the oxidative switch of angiotensinogen to its more active sulphydryl-bridged form in the maternal circulation in pre-eclampsia-the hypertensive crisis of pregnancy that threatens the health and survival of both mother and child

Structural biology of two proteins from two eye-related protein families : ABCR and TSP36

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The NO donor sodium nitroprusside: evaluation of skeletal muscle vascular and metabolic dysfunction

The nitric oxide (NO) donor sodium nitroprusside (SNP) may promote cyanide-induced toxicity and systemic and/or local responses approaching maximal vasodilation. The hypotheses were tested that SNP superfusion of the rat spinotrapezius muscle exerts 1) residual impairments in resting and contracting blood flow, oxygen utilization (VO[subscript 2]) and microvascular O[subscript 2] pressure (PO[subscript 2mv]); and 2) marked hypotension and elevation in resting PO[subscript 2mv]. Two superfusion protocols were performed: 1) Krebs-Henseleit (control 1), SNP (300 μM; a dose used commonly in superfusion studies) and Krebs-Henseleit (control 2), in this order; 2) 300 and 1200 μM SNP in random order. Spinotrapezius muscle blood flow (radiolabeled microspheres), VO[subscript 2] (Fick calculation) and PO[subscript 2mv] (phosphorescence quenching) were determined at rest and during electrically-induced (1 Hz) contractions. There were no differences in spinotrapezius blood flow, VO[subscript 2] or PO[subscript 2mv] at rest and during contractions pre- and post-SNP condition (control 1 and control 2; p>0.05 for all). With regard to dosing, SNP produced a graded elevation in resting PO[subscript 2mv] (p<0.05) with a reduction in mean arterial pressure only at the higher concentration (p<0.05). Contrary to our hypothesis, skeletal muscle superfusion with the NO donor SNP (300 μM) improved microvascular oxygenation during the transition from rest to contractions (PO[subscript 2mv] kinetics) without precipitating residual impairment of muscle hemodynamic or metabolic control or compromising systemic hemodynamics. These data suggest that SNP superfusion (300 μM) constitutes a valid and important tool for assessing the functional roles of NO in resting and contracting skeletal muscle function without incurring residual alterations consistent with cyanide accumulation and poisoning

Prolonged Exposure to Monosodium Glutamate in Healthy Young Adults Decreases Perceived Umami Taste and Diminishes Appetite for Savory Foods

Background; Research suggests that increased consumption of sweet, salt, or fat is associated with diminished perceived taste intensity and shifted preferences for the respective stimulus. It is unknown whether a similar effect occurs with the consumption of umami. Objective; The aim of the study was to investigate the influence of habitual exposure to umami stimuli on umami taste perception, hedonics, and satiety. Methods; Fifty-eight healthy men (n = 16) and women (n = 42) participated in a parallel-group, randomized controlled study. The normal-weight [mean ± SD body mass index (kg/m2): 21.8 ± 2.2] group of young adults (mean ± SD age: 22.7 ± 6.2 y) consumed vegetable broth daily for 4 wk. The broth for the treatment group (n = 28) was supplemented with 3.8 g monosodium glutamate (MSG), whereas the control group (n = 30) consumed a sodium-matched broth without MSG. Perceived umami taste intensity and discrimination in MSG solutions; liking, wanting, and preference of a variety of umami-rich foods; satiation and satiety from an ad libitum meal; and anthropometric measures were evaluated at baseline and at week 4. General linear models assessed the effect of treatment on change from baseline for all outcomes and tested for effect modification of sex. Results; Relative to controls, increased consumption of MSG for 4 wk diminished umami taste in women (8.4 units on generalized Labeled Magnitude Scale; 95% CI: –13.8, –3.1 units; P = 0.013). The desire for and intake of savory foods decreased after MSG treatment in both sexes with an ad libitum meal (desire: –7.7 units; 95% CI: –13.7, –1.7 units; P = 0.04; intake: –36 g; 95% CI: –91, 19 g; P = 0.04). Conclusion; Our results highlight that a month-long diet high in umami stimuli attenuates perceived umami taste and appetite for savory foods in a young, healthy population. Our findings contribute to the understanding of food choice, a factor in the development and maintenance of obesity, as well as the etiology of protein-related health conditions such as osteoporosis and kidney disease. This study is registered at www.clinicaltrials.gov as NCT03010930