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Proceedings of the National Academy of... Jul 2023The serotonin transporter (SERT) is a member of the SLC6 neurotransmitter transporter family that mediates serotonin reuptake at presynaptic nerve terminals. SERT is the...
The serotonin transporter (SERT) is a member of the SLC6 neurotransmitter transporter family that mediates serotonin reuptake at presynaptic nerve terminals. SERT is the target of both therapeutic antidepressant drugs and psychostimulant substances such as cocaine and methamphetamines, which are small molecules that perturb normal serotonergic transmission by interfering with serotonin transport. Despite decades of studies, important functional aspects of SERT such as the oligomerization state of native SERT and its interactions with potential proteins remain unresolved. Here, we develop methods to isolate SERT from porcine brain (pSERT) using a mild, nonionic detergent, utilize fluorescence-detection size-exclusion chromatography to investigate its oligomerization state and interactions with other proteins, and employ single-particle cryo-electron microscopy to elucidate the structures of pSERT in complexes with methamphetamine or cocaine, providing structural insights into psychostimulant recognition and accompanying pSERT conformations. Methamphetamine and cocaine both bind to the central site, stabilizing the transporter in an outward open conformation. We also identify densities attributable to multiple cholesterol or cholesteryl hemisuccinate (CHS) molecules, as well as to a detergent molecule bound to the pSERT allosteric site. Under our conditions of isolation, we find that pSERT is best described as a monomeric entity, isolated without interacting proteins, and is ensconced by multiple cholesterol or CHS molecules.
Topics: Animals; Swine; Serotonin Plasma Membrane Transport Proteins; Cryoelectron Microscopy; Detergents; Serotonin; Central Nervous System Stimulants; Cocaine; Methamphetamine
PubMed: 37436958
DOI: 10.1073/pnas.2304602120 -
Animals : An Open Access Journal From... Aug 2023The German Society of Nutrition Physiology has proposed a new protein evaluation system for horse feeds to estimate pre-cecally digestible crude protein (pcdCP) and...
The German Society of Nutrition Physiology has proposed a new protein evaluation system for horse feeds to estimate pre-cecally digestible crude protein (pcdCP) and amino acids (pcdAA) from chemical properties. A total of 71 feeds for horses were chemically tested and evaluated according to the new protein evaluation system. A feeding trial with eight horses tested whether differences in estimated pcdAA and neutral detergent soluble CP (NDSCP) in the diet were reflected by post-prandial (ppr) kinetics of plasma lysine (Lys) by feeding a complementary feed (control = CTRL) with 1.02 g Lys/100 kg body weight (BW) as well as three diets with 3.02 g Lys/100 kg BW, as follows: (i) CTRL with synthetic AA (CTRL + synAA); (ii) CTRL with soybean meal (CTRL + SBM); and (iii) lucerne pellets (LUC). In comparison to CTRL, the areas of curves (s) of ppr plasma Lys differed: CTRL < CTRL + SBM ( < 0.01) < CTRL + synAA ( < 0.05). For 71 feeds, the estimated pcdCP was correlated with the CP content ( < 0.001), NDSCP ( < 0.001), and ash-free neutral detergent fiber ( < 0.001). A mean neutral detergent insoluble CP content of at least 3-5% can be assumed in horse feed. It is speculated that the predicted availability of Lys from LUC seems to be underestimated by the new protein evaluating system. The influence of chewing and microbiota in vivo needs to be considered in horses.
PubMed: 37627415
DOI: 10.3390/ani13162624 -
Biochemical Society Transactions Jun 2023Membrane proteins are a highly relevant class of biological molecules and comprise ∼60% of current drug targets. Before being analyzed by structural, biochemical, and... (Review)
Review
Membrane proteins are a highly relevant class of biological molecules and comprise ∼60% of current drug targets. Before being analyzed by structural, biochemical, and biophysical methods, membrane proteins must first be extracted from cellular membranes - often using detergents. Detergent-extracted membrane proteins are amenable to analysis by structural, biochemical, and biophysical techniques. In certain cases, however, detergents can disturb native protein conformations and/or biological activity. This has led to the development of membrane mimetics, which stabilize membrane proteins in a native membrane-like environment that is water-soluble and detergent-free. This review provides an overview of recent developments in the membrane mimetic field, with a focus on nanodiscs, Saposin lipid nanoparticles (SapNPs), peptidiscs, and SMA lipid particles (SMALPs) - and highlights their utility for supporting biophysical, biochemical, and structural characterization of membrane proteins and complexes.
Topics: Membrane Proteins; Cell Membrane; Biomimetics; Lipid Bilayers; Detergents
PubMed: 37345653
DOI: 10.1042/BST20230164 -
Journal of Cachexia, Sarcopenia and... Dec 2023Sarcopenia in liver cirrhosis is associated with low quality of life and high mortality risk. The pathogenesis has yet to be fully understood. We hypothesized that gut...
BACKGROUND
Sarcopenia in liver cirrhosis is associated with low quality of life and high mortality risk. The pathogenesis has yet to be fully understood. We hypothesized that gut microbiome, bile acid (BA) composition and metabolites differ between cirrhotic patients with and without sarcopenia and contribute to pathogenesis.
METHODS
Cirrhotic patients with (n = 78) and without (n = 38) sarcopenia and non-cirrhotic controls with (n = 39) and without (n = 20) sarcopenia were included in this study. Faecal microbiome composition was studied by 16S rDNA sequencing, serum and faecal BA composition by ultra-high-performance liquid chromatography-tandem mass spectrometry, and metabolite composition in serum, faeces and urine by nuclear magnetic resonance.
RESULTS
Bacteroides fragilis, Blautia marseille, Sutterella spp. and Veillonella parvula were associated with cirrhotic patients with sarcopenia, whereas Bacteroides ovatus was more abundant in cirrhotic patients without sarcopenia. We observed significantly elevated secondary BAs, deoxycholic acid (DCA; P = 0.01) and lithocholic acid (LCA; P = 0.02), and the ratios of deoxycholic acid to cholic acid (DCA:CA; P = 0.04), lithocholic acid to chenodeoxycholic acid (LCA:CDCA; P = 0.03) and 12 alpha-hydroxylated to non-12 alpha-hydroxylated BAs (12-α-OH:non-12-α-OH BAs; P = 0.04) in serum of cirrhotic patients with sarcopenia compared with cirrhotic patients without sarcopenia, indicating an enhanced transformation of primary to secondary BAs by the gut microbiome. CA (P = 0.02) and the ratios of CA:CDCA (P = 0.03) and total ursodeoxycholic acid to total secondary BAs (T-UDCA:total-sec-BAs, P = 0.03) were significantly reduced in the stool of cirrhotic patients with sarcopenia compared with cirrhotic patients without sarcopenia. Also, valine and acetate were significantly reduced in the serum of cirrhotic patients with sarcopenia compared with cirrhotic patients without sarcopenia (P = 0.01 and P = 0.03, respectively). Multivariate logistic regression further confirmed the association of B. ovatus (P = 0.01, odds ratio [OR]: 12.8, 95% confidence interval [CI]: 168.1; 2.2), the ratios of 12-α-OH:non-12-α-OH BAs (P = 0.03, OR: 2.54, 95% CI: 0.99; 6.55) and T-UDCA:total-sec-BAs (P = 0.04, OR: 0.25, 95% CI: 0.06; 0.98) in serum and stool CA:CDCA (P = 0.04, OR: 0.79, 95% CI: 0.62; 0.99), and serum valine (P = 0.04, OR: 1.00, 95% CI: 1.02; 1.00) with sarcopenia in cirrhosis after correcting for the severity of liver disease and sex.
CONCLUSIONS
Our study suggests a potential functional gut microbiome-host interaction linking sarcopenia with the altered gut microbiomes, BA profiles and amino acids pointing towards a potential mechanistic interplay in understanding sarcopenia pathogenesis.
Topics: Humans; Bile Acids and Salts; Gastrointestinal Microbiome; Quality of Life; Sarcopenia; Liver Cirrhosis; Lithocholic Acid; Metabolome; Deoxycholic Acid; Valine
PubMed: 37767786
DOI: 10.1002/jcsm.13342 -
Nature Communications Aug 2023Hydroxysteroid 17-beta-dehydrogenase 13 (HSD17B13) is a hepatic lipid droplet-associated enzyme that is upregulated in patients with non-alcoholic fatty liver disease....
Hydroxysteroid 17-beta-dehydrogenase 13 (HSD17B13) is a hepatic lipid droplet-associated enzyme that is upregulated in patients with non-alcoholic fatty liver disease. Recently, there have been several reports that predicted loss of function variants in HSD17B13 protect against the progression of steatosis to non-alcoholic steatohepatitis with fibrosis and hepatocellular carcinoma. Here we report crystal structures of full length HSD17B13 in complex with its NAD cofactor, and with lipid/detergent molecules and small molecule inhibitors from two distinct series in the ligand binding pocket. These structures provide insights into a mechanism for lipid droplet-associated proteins anchoring to membranes as well as a basis for HSD17B13 variants disrupting function. Two series of inhibitors interact with the active site residues and the bound cofactor similarly, yet they occupy different paths leading to the active site. These structures provide ideas for structure-based design of inhibitors that may be used in the treatment of liver disease.
Topics: Humans; Carcinoma, Hepatocellular; Lipid Droplet Associated Proteins; Lipids; Liver Neoplasms; Non-alcoholic Fatty Liver Disease; 17-Hydroxysteroid Dehydrogenases
PubMed: 37620305
DOI: 10.1038/s41467-023-40766-0 -
ACS Applied Materials & Interfaces Jun 2023Hydrophilic/oleophobic surfaces are desirable in many applications including self-cleaning, antifogging, oil-water separation, etc. However, making plastic surfaces...
Hydrophilic/oleophobic surfaces are desirable in many applications including self-cleaning, antifogging, oil-water separation, etc. However, making plastic surfaces hydrophilic/oleophobic is challenging due to the intrinsic hydrophobicity/oleophilicity of plastics. Here, we report a simple and effective method of making plastics hydrophilic/oleophobic. Plastics, including poly (methyl methacrylate) (PMMA), polystyrene (PS), and polycarbonate (PC), have been coated with a perfluoropolyether (PFPE) (i.e., commercially known as Zdol) via dip coating and then irradiated with UV/Ozone. The contact angle measurements indicate that the treated plastics have a lower water contact angle (WCA) and higher hexadecane contact angle (HCA), i.e., they are simultaneously hydrophilic/oleophobic. The Fourier transform infrared (FTIR) results suggest that UV/Ozone treatment introduces oxygen-containing polar groups on the plastic surfaces, which renders the plastic surfaces hydrophilic. Meanwhile, more orderly packed PFPE Zdol molecules, which is due to the UV-induced bonding between PFPE Zdol and the plastic surface, result in the oleophobicity. Moreover, the simultaneous hydrophilicity/oleophobicity of functionalized plastics does not degrade in aging tests, and they have superior antifogging performance and detergent-free cleaning capability. This simple method developed here potentially can be applied to other plastics and has important implications in the functionalization of plastic surfaces.
PubMed: 37326374
DOI: 10.1021/acsami.3c06787 -
MethodsX Jun 2024Surfactants are indispensable in industrial applications today due to their wetting, emulsifying, dispersing, cleansing, and detergent properties. The use of surfactants...
Surfactants are indispensable in industrial applications today due to their wetting, emulsifying, dispersing, cleansing, and detergent properties. The use of surfactants extends from the cosmetic industry to the petroleum industry and beyond. Their characteristics and effectiveness can be assessed through various standardized tests, and based on these methods, their applications can be determined. However, there is a lack of a universally applicable testing method for one crucial and complex property: the detergent effect. The detergent effect refers to the removal of unwanted contaminants from a solid surface. However, cleaning is not solely attributed to the surfactant but to the appropriate combination of various factors, whose synergistic effect reduces surface contamination. The most significant factors influencing detergent effect include the characteristics and nature of the contaminants, properties of the cleaning solution (surfactant concentration and composition, water hardness, enzymes, etc.), temperature, washing time, and hydrodynamic conditions. Additionally, the presence of electrolytes, pH of the cleaning solution, and detergent foaming properties may also play important roles. Our goal was to develop a detergent effect testing methodology that is not specific to any particular application domain but offers a straightforward and easy-to-implement solution for comparing the detergent effect of various types of surfactants.•The study presents a method for determining detergent effect of surfactants.•The method is universal and suitable for the evaluation of any type of surfactant.•The method is low-cost and easy to perform.
PubMed: 38799038
DOI: 10.1016/j.mex.2024.102755 -
Clinical Drug Investigation Dec 2023Phase II studies on tauro-urso-deoxycholic acid (TUDCA) raised the promise of safety and efficacy in patients with amyotrophic lateral sclerosis, a currently incurable... (Review)
Review
Phase II studies on tauro-urso-deoxycholic acid (TUDCA) raised the promise of safety and efficacy in patients with amyotrophic lateral sclerosis, a currently incurable and devastating disease. We review the available evidence on the efficacy and safety of TUDCA, administered alone or in combination, by analyzing and comparing published and ongoing studies on amyotrophic lateral sclerosis. Two independent phase II studies (using TUDCA solo or combined with sodium phenylbutyrate) showed similar efficacy in slowing disease progression measured by functional scales. One open-label follow-up TUDCA+sodium phenylbutyrate study suggested a benefit on survival. Two subsequent phase III studies with TUDCA (solo or combined with sodium phenylbutyrate) have been initiated and are currently ongoing. Their completion is expected by the end of 2023 and beginning of 2024. Evidence collected by phase II studies indicates that there are no safety concerns in patients with amyotrophic lateral sclerosis. The efficacy shown in phase II studies was considered sufficient to grant approval in some countries but not in others, owing to discrepant views on the strength of evidence. It will be necessary to wait for the results of ongoing phase III studies to attain a full appreciation of these data.
Topics: Humans; Amyotrophic Lateral Sclerosis; Phenylbutyrates; Taurochenodeoxycholic Acid
PubMed: 37973672
DOI: 10.1007/s40261-023-01324-0 -
Journal of Colloid and Interface Science Feb 2024Understanding the complex interactions between polymers and surfactants is required to optimise commercially relevant systems such as paint, toothpaste and detergent....
HYPOTHESIS
Understanding the complex interactions between polymers and surfactants is required to optimise commercially relevant systems such as paint, toothpaste and detergent. Neutral polymers complex with surfactants, forming 'pearl necklace' structures that are often conceptualised as pseudo-polyelectrolytes. Here we pose two questions to test the limits of this analogy: Firstly, in the presence of salt, do these polymer-surfactant systems behave like polyelectrolytes? Secondly, do polymer-surfactant complexes resist geometric confinement like polyelectrolytes?
EXPERIMENTS
We test the limits of the pseudo-polyelectrolyte analogy through studying a poly(N-isopropylacrylamide) (PNIPAM) brush in the presence of sodium dodecylsulfate (SDS). Brushes are ideal for interrogating pseudo-polyelectrolytes, as neutral and polyelectrolyte brushes exhibit distinct and well understood behaviours. Spectroscopic ellipsometry, quartz crystal microbalance with dissipation monitoring (QCM-D), and neutron reflectometry (NR) were used to monitor the behaviour and structure of the PNIPAM-SDS system as a function of NaCl concentration. The ability of the PNIPAM-SDS complex to resist geometric confinement was probed with NR.
FINDINGS
At a fixed SDS concentration below the zero-salt CMC, increasing NaCl concentration <100 mM promoted brush swelling due to an increase in osmotic pressure, not dissimilar to a weak polyelectrolyte. At these salt concentrations, the swelling of the brush could be described by a single parameter: the effective CMC. However, at high NaCl concentrations (e.g., 500 mM) no brush collapse was observed at all (non-zero) concentrations of SDS studied, contrary to what is seen for many polyelectrolytes. Study of the polymer-surfactant system under confinement revealed that the physical volume of surfactant dominates the structure of the strongly confined system, which further differentiates it from the polyelectrolyte case.
PubMed: 37944374
DOI: 10.1016/j.jcis.2023.10.101