Lipid Mobilization Research Articles

Fluorescence Correlation Spectroscopy Reveals Interaction of Some Microdomain-Associated Lipids with Cellular Focal Adhesion Sites.

Cells adhere to the extracellular matrix at distinct anchoring points, mostly focal adhesions. These are rich in immobile transmembrane- and cytoskeletal-associated proteins, some of which are known to interact with lipids of the plasma membrane. To investigate their effect on lipid mobility and molecular interactions, fluorescently labeled lipids were incorporated into the plasma membranes of primary myofibroblasts using fusogenic liposomes. With fluorescence correlation spectroscopy, we tested mobilities of labeled microdomain-associated lipids such as sphingomyelin (SM), ganglioside (GM1), and cholesterol as well as of a microdomain-excluded phospholipid (PC) and a lipid-like molecule (DiIC18(7)) in focal adhesions (FAs) and in neighboring non-adherent membrane areas. We found significantly slower diffusion of SM and GM1 inside FAs but no effect on cholesterol, PC, and DiIC18(7). These data were compared to the molecular behavior in Lo/Ld-phase separated giant unilamellar vesicles, which served as a model system for microdomain containing lipid membranes. In contrast to the model system, lipid mobility changes in FAs were molecularly selective, and no particle enrichment occurred. Our findings suggest that lipid behavior in FAs cannot be described by Lo/Ld-phase separation. The observed slow-down of some molecules in FAs is potentially due to transient binding between lipids and some molecular constituent(s).

Phenolic Compounds Reduce the Fat Content in Caenorhabditis elegans by Affecting Lipogenesis, Lipolysis, and Different Stress Responses.

Supplementation with bioactive compounds capable of regulating energy homeostasis is a promising strategy to manage obesity. Here, we have screened the ability of different phenolic compounds (myricetin, kaempferol, naringin, hesperidin, apigenin, luteolin, resveratrol, curcumin, and epicatechin) and phenolic acids (p-coumaric, ellagic, ferulic, gallic, and vanillic acids) regulating C. elegans fat accumulation. Resveratrol exhibited the strongest lipid-reducing activity, which was accompanied by the improvement of lifespan, oxidative stress, and aging, without affecting worm development. Whole-genome expression microarrays demonstrated that resveratrol affected fat mobilization, fatty acid metabolism, and unfolded protein response of the endoplasmic reticulum (UPRER), mimicking the response to calorie restriction. Apigenin induced the oxidative stress response and lipid mobilization, while vanillic acid affected the unfolded-protein response in ER. In summary, our data demonstrates that phenolic compounds exert a lipid-reducing activity in C. elegans through different biological processes and signaling pathways, including those related with lipid mobilization and fatty acid metabolism, oxidative stress, aging, and UPR-ER response. These findings open the door to the possibility of combining them in order to achieve complementary activity against obesity-related disorders.

Whole-body vibration promotes lipid mobilization in hypothalamic obesity rat

ObjectivesThis study aimed to analyze the effect of whole-body vibration (WBV) on metabolic parameters using the monosodium l-glutamate (MSG) model of obesity. MethodMSG-obese rats that were exposed to WBV on a vibrating platform with 60 Hz frequency, 2 mm amplitude, three times/week, 10 min/day, during eight weeks (from postnatal day (PN) 80 to PN136). Blood glucose, creatine kinases (CK and CK-MB) and lipid profile through plasma and liver levels of lipids and lipoproteins were evaluated. Morphology and oxidative stress of adipose and hepatic tissues were further evaluated. ResultsWhen performing a WBV exercise, animals showed contrasting metabolic responses. Vibration Control group (CTL-WBV) presented a reduction in CK and liver triacylglycerol, an increase in glucose, lactate, total cholesterol, liver cholesterol, and LDL while MSG Vibration group (MSG-WBV) showed an increase in total triacylglycerol, VLDL, lactate, CK, liver cholesterol, additional liver lipid peroxidation and LDL, total cholesterol and CKMB reduction. ConclusionEven although the MSG is a model of impacting injury, the metabolic demand of WBV exercise was able to induce mobilization of substrates, highlighting the lipid mobilization in obese animals, it should be used as a metabolic rehabilitation tool in patients with metabolic diseases, such as obesity and diabetes.

Molecular Dynamics Study of Lipid and Cholesterol Reorganization Due to Membrane Binding and Pore Formation by Listeriolysin O.

Pore-forming toxins are proteinsexpressed by bacteria to primarily cause infections in the host cell. Cholesterol-dependent cytolysins (CDCs) are a class of proteins whose pore-forming ability requires the presence of cholesterol in the membrane. Upon binding to the target cell, cholesterol-recognizing residues in the membrane binding D4 subdomain assist in stabilizing both the pre-pore and pore states which occur during protein oligomerization on the cell membrane. Super resolution-stimulated emission depletion (STED) microscopy experiments (Sarangi et al. in Langmuir, 32:9649-9657, 2016) on supported lipid bilayers have shown that listeriolysin (LLO), a CDC expressed by Listeria monocytogenes, a food-borne pathogen, induces both spatial and dynamic heterogeneity in bilayer membranes. Here, we use all-atom molecular dynamics simulations to explore molecular details of the induced membrane reorganization by considering two distinct states of the oligomerized LLO protein in a 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC): cholesterol membrane. In the membrane bound (MB) state, four D4 subunits are placed at the bilayer interface in a pre-pore configuration and the membrane-inserted (MI) state consists of a tetrameric arc-like pore configuration. By analyzing lipid-order parameters, mobilities, and diffusion coefficients, we examine the induced spatial heterogeneity that occurs in both the MB and MI states. This heterogeneity is primarily driven by the local density enhancement of cholesterol in the vicinity of the MB, D4 subunits leading to distinct differences in lipid and cholesterol mobility across the two leaflets as well as enhanced lipid mobilities in regions where cholesterol is depleted. The leaflet-induced heterogeneity is greater for the MB state when compared with the MI state and the dynamic variations are more pronounced in the extracellular leaflet when compared with the cytosolic leaflet. Our study provides molecular-level insights into the inhomogeneity and perturbation induced in bilayer membranes upon LLO binding and pore formation and is expected to represent trends across PFTs in the broad CDC subclass of proteins.

Single-molecule analysis of dynamics and interactions of the SecYEG translocon.

Protein translocation and insertion into the bacterial cytoplasmic membrane are the essential processes mediated by the Sec machinery. The core machinery is composed of the membrane-embedded translocon SecYEG that interacts with the secretion-dedicated ATPase SecA and translating ribosomes. Despite the simplicity and the available structural insights on the system, diverse molecular mechanisms and functional dynamics have been proposed. Here, we employ total internal reflection fluorescence microscopy to study the oligomeric state and diffusion of SecYEG translocons in supported lipid bilayers at the single-molecule level. Silane-based coating ensured the mobility of lipids and reconstituted translocons within the bilayer. Brightness analysis suggested that approx. 70% of the translocons were monomeric. The translocons remained in a monomeric form upon ribosome binding, but partial oligomerization occurred in the presence of nucleotide-free SecA. Individual trajectories of SecYEG in the lipid bilayer revealed dynamic heterogeneity of diffusion, as translocons commonly switched between slow and fast mobility modes with corresponding diffusion coefficients of 0.03 and 0.7µm2 ·s-1 . Interactions with SecA ATPase had a minor effect on the lateral mobility, while bound ribosome:nascent chain complexes substantially hindered the diffusion of single translocons. Notably, the mobility of the translocon:ribosome complexes was not affected by the solvent viscosity or macromolecular crowding modulated by Ficoll PM 70, so it was largely determined by interactions within the lipid bilayer and at the interface. We suggest that the complex mobility of SecYEG arises from the conformational dynamics of the translocon and protein:lipid interactions.

Mobilization of storage lipid reserve and expression analysis of lipase and lipoxygenase genes in rice (Oryza sativa var. Pusa Basmati 1) bran during germination

Storage lipid mobilization by lipases and lipoxygenases (LOXs) in response to developmental cues take place during seed germination. After rice grain milling, the endogenous lipases and LOXs present in the bran fraction come in contact with the storage lipid reserve or triacylglycerol (TAG). Lipases catalyze the hydrolysis of TAGs to non-esterified fatty acids (NEFAs) and glycerol. The NEFAs, especially linoleic acid (18:2) produced, are further subjected to oxidative rancidity via peroxidation reaction catalyzed by LOXs. This results in the production of conjugated hydroperoxides of 18:2 that influence the off-flavors in rice bran lipids. The aim of this study is to understand how lipid mobilization and expression of lipase and LOX genes occur in the bran of germinating rice grains (Oryza sativavar. Pusa Basmati 1). Our results show that the primary source of storage lipids in bran is TAG, and its mobilization starts at 4 days after imbibition (4 DAI). Using publically available RNA-seq data and phylogeny analyses, we selected a total of 18 lipase and 16 LOX genes in rice for their expression profiles during onset of lipid mobilization. Gene expression analyses revealed OsLip1, OsLip9, and OsLip13; and OsLOX3 and OsLOX14 as the predominantly expressed genes in bran of germinating rice grains. This study explores two important events in the germinating rice grains, namely, mobilization of storage lipids and expression pattern of lipase and LOX genes. The information generated in this study can be used to efficiently manipulate the genes to enhance the shelf-stability of bran lipid reserve.

Cardiotrophin-1 contributes to metabolic adaptations through the regulation of lipid metabolism and to the fasting-induced fatty acid mobilization.

It is becoming clear that several human pathologies are caused by altered metabolic adaptations. During liver development, there are physiological changes, from the predominant utilization of glucose (fetal life) to the use of lipids (postnatal life). Fasting is another physiological stress that elicits well-known metabolic adjustments. We have reported the metabolic properties of cardiotrophin-1 (CT-1), a member of the interleukin-6 family of cytokines. Here, we aimed at analyzing the role of CT-1 in response to these metabolic changes. We used different in vivo models. Furthermore, a differential study was carried out with wild-type and CT-1 null mice in fed (ad libitum) and food-restricted conditions. We demonstrated that Ct-1 is a metabolic gene induced in the liver via PPARα in response to lipids in mice (neonates- and food-restricted adults). We found that Ct-1 mRNA expression in white adipose tissue directly involved PPARα and PPARγ. Finally, the physiological role of CT-1 in fasting is confirmed by the impaired food restriction-induced adipose tissue lipid mobilization in CT-1 null mice. Our findings support a previously unrecognized physiological role of CT-1 in metabolic adaptations, through the regulation of lipid metabolism and contributes to fasting-induced free fatty acid mobilization.

Effect of Vesicle Size on the Cytolysis of Cell-Penetrating Peptides (CPPs).

A specific series of peptides, called a cell-penetrating peptide (CPP), is known to be free to directly permeate through cell membranes into the cytosol (cytolysis); hence, this CPP would be a potent carrier for a drug delivery system (DDS). Previously, we proposed the mechanism of cytolysis as a temporal and local phase transfer of membrane lipid caused by positive membrane curvature generation. Moreover, we showed how to control the CPP cytolysis. Here, we investigate the phospholipid vesicle’s size effect on CPP cytolysis because this is the most straightforward way to control membrane curvature. Contrary to our expectation, we found that the smaller the vesicle diameter (meaning a higher membrane curvature), the more cytolysis was suppressed. Such controversial findings led us to seek the reason for the unexpected results, and we ended up finding out that the mobility of membrane lipids as a liquid crystal is the key to cytolysis. As a result, we could explain the cause of cytolysis suppression by reducing the vesicle size (because of the restriction of lipid mobility); osmotic pressure reduction to enhance positive curvature generation works as long as the membrane is mobile enough to modulate the local structure. Taking all the revealed vital factors and their effects as a tool, we will further explore how to control CPP cytolysis for developing a DDS system combined with appropriate cargo selection to be tagged with CPPs.

Do biological and management reasons for a short or long dry period induce the same effects on dairy cattle productivity?

The dry period is a well-established factor that determines lactation success. A retrospective observational study used 32,182 lactations from 16 farms to determine whether management versus biological reasons for deviations from the targeted 60-d dry period have the same associations with subsequent lactation performance. Herd inclusion criteria were Holstein cows, herd size ≥900 cows, breeding by artificial insemination, and (minimally) bimonthly milk testing. Dry period length (DPL) and gestation length (GL) were each categorized as short [>1 standard deviation (SD) below mean within herd; means 45 d DPL, 269 d GL] or long (>1 SD above mean within herd; means 73 d DPL, 284 d GL) and combined to generate the following 7 study groups: short DPL, short GL (SDSG, n = 2,123); short DPL, average GL (SDAG, n = 1,418); average DPL, short GL (ADSG, n = 1,759); average DPL, average GL (ADAG, n = 19,265); average DPL, long GL (ADLG, n = 3,325); long DPL, average GL (LDAG, n = 2,573); and long DPL, long GL (LDLG, n = 1,719). Responses evaluated included milk and component yields at first test and over the whole lactation, days to first service, first service conception risk, days open, and herd retention through 60 and 365 d postpartum. Continuous data were analyzed by mixed models and time to event data by Cox proportional hazard models, both accounting for clustering at the herd level. First test and whole-lactation milk and component yields were lowest for SDSG. Within cows that experienced calving difficulty, rates of receiving first service were 13 and 20% less for SDSG and ADSG compared with ADAG. Hazard of leaving the herd by 60 d in milk (DIM) was 34% greater for ADSG than ADAG. Similar effects between SDSG and ADSG but not SDAG indicated that short GL was a greater contributor to poor performance than DPL itself. Overall production was similar between ADAG and SDAG; however, somatic cell linear score at first test was greater for SDAG, and milk yield at first test was lesser for SDAG cows with greater milk at last test before dry-off. Although short DPL might be a successful strategy for some herds or cows, cows with high milk yield at dry-off should not be subjected to a short dry period. Long DPL or GL did not influence early-lactation or whole-lactation milk yield. Cows with a long DPL due to early dry-off (LDAG) likely experienced issues related to excessive lipid mobilization, as milk fat concentration and fat:protein ratio at first test were greater and hazard of leaving the herd was 30 and 24% greater compared with ADAG by 60 and 365 DIM, respectively. We conclude that deviations in DPL length caused by biology (short GL) were associated with greater effects than management causes of short DPL, whereas management reasons for long DPL were associated with more negative outcomes than long GL.

Assessment of Growth, Lipid Metabolism and Gene Expression Responses in Senegalese Sole Larvae Fed With Low Dietary Phospholipid Levels.

Phospholipids (PL) are essential molecules for larval growth and development. In this study, growth, lipid metabolism and gene expression responses associated with different dietary PL levels in pelagic sole larvae were evaluated. In a first trial, the long-term effects on growth and survival of two experimental microdiets (MD) containing high (High-PL) or low (Low-PL) PL levels were tested and compared to a diet based on live prey (rotifers). The MD were supplied from 3 to 10 days post-hatch (dph) and Artemia from day 8 to 29 dph. High-PL fed larvae had higher dry mass (1.2-fold) than Low-PL fed larvae at 8 dph and both MD were smaller (2.9-fold) than larvae fed live preys. However, a compensatory growth (33% between 8 and 20 dph) occurred when MD were substituted by Artemia and by the end of the trial no significant differences in mass or survival occurred between the dietary treatments. In a second trial, growth, lipid metabolism and gene expression profiles of larvae fed with MD up to 8 dph were analyzed. Growth data confirmed that mass of larvae fed with High-PL was higher (1.3-fold) than the those fed Low-PL and they had lower levels of triacylglycerol (2.8-fold), cholesterol (1.2-fold) and cetoleic acid (1.7-fold). Histological analysis indicated an excess of lipid vacuoles in larvae fed with Low-PL and the expression analysis revealed a coordinated response to enhance lipid mobilization since the expression of genes involved in PL intermediate synthesis, PL remodeling as well as eight apolipoprotein was up-regulated. The down-regulation of apolipoprotein apob2 in larvae fed with Low-PL indicated a specific regulation by PL levels. The present work provides insight into the responses associated with dietary PL in early fish larvae, which will be of use for future studies aimed as designing effective larval sole diets.

Characterization of serum adiponectin and leptin in healthy perinatal dairy cows or cows with ketosis, and their effectson ketosis involved indices.

We investigated changes in concentrations of ADP (adiponectin), LEP (leptin), BHBA (beta-hydroxybutyric acid), NEFA (non-esterified fatty acid), Glucose (Glu) and INS (insulin) in serum of healthy perinatal dairy cows and cows with ketosis. Twenty-one healthy cows and seventeen cows with ketosis from a herd of a total 60 Holstein cows (near dry period i.e. 56 days antepartum) were selected. Blood was collected through the tail vein every 7 days, from 56 day antepartum to 56 day postpartum. Serum ADP, LEP, BHBA, NEFA, Glu, and INS concentrations were determined, and ketosis was diagnosed through serum BHBA (≥1.2 mmol/L). We showed the concentration of serum adipokines and energy balancing indices were stable during antepar- tum period. However, ADP concentration increased while LEP decreased, and there were a significant increase in cows with ketosis compared to that of in healthy cows. Serum BHBA and NEFA concentrations increased significantly at first, and then gradually decreased in both healthy cows and cows with ketosis. However, cows with ketosis showed higher concentrations of BHBA and NEFA which restored later. The serum concentration of Glu in both healthy dairy cows and cows with ketosis showed a decreasing trend. INS concentration in healthy cows was decreased while it was increased in cows with ketosis. The results reflect the extent of hypo- glycemia and lipid mobilization postpartum, suggest IR exists in cows with ketosis while serum ADP and LEP might play roles in the development of ketosis.

Heterogeneous Differentiation of Highly Proliferative Embryonal Carcinoma PCC4 Cells Induced by Curcumin: An In Vitro Study

Curcumin, the yellow pigment derived from turmeric rhizomes, exhibits antioxidant, anti-inflammatory, antimicrobial, and anticancer properties. We have previously reported in a study that curcumin could induce differentiation in embryonal carcinoma cell (EC). EC cells are the primary constituents of teratocarcinoma tumors, and hence differentiating them to a non-proliferative cell type may be useful in anticancer therapies. Here, we conducted a detailed study using various molecular approaches to characterize this differentiation at the cellular and molecular levels. The cells were treated with 20 µM curcumin, which was the optimal concentration to produce the highest amount of differentiated cells. Changes in protein and RNA expression, membrane dynamics, and migration of these cells after treatment with curcumin were then studied in a time-dependent manner. The differentiated cells were morphologically distinct from the precursor cells, and gene expression profiles were altered in curcumin-treated cells. Curcumin promoted cell motility and cell adhesion. Curcumin also induced changes in membrane fluidity and the lateral mobility of lipids in the plasma membrane. The findings of this study suggest that curcumin might have therapeutic potential in differentiation therapy for the treatment of teratocarcinomas or germ cell tumors (GCTs) such as testicular and ovarian GCTs.

Regulation of Properties of Lipid Membranes by Interaction with 2-Hydroxypropyl β-Cyclodextrin: Molecular Details

The interaction between 2-hydroxypropyl β-cyclodextrin (HPCD) and a liposomal bilayer has been studied. The main binding sites of HPCD on the surface of neutral liposomes based on dipalmitoylphosphatidylcholine (DPPC) are the phosphate groups of lipids. The complex formation with HPСD leads to the stabilization of the gel state of monocomponent liposomes. The inclusion of the anionic component, cardiolipin (CL, 20%), in the bilayer changes the nature of the liposome-HPCD interaction. The observed lipid disorder disturbs membrane integrity, which is revealed in the release of the included dye (phenolphthalein). The effect of HPCD on the process and parameters of the phase transition of anionic liposomes has been studied using thermograms, which show the change in the position of the absorption bands of lipid acyl chains in the FTIR spectrum of liposomes. A stratification of DPPC/CL (80/20%) bilayer into two microphases with different cardiolipin content has been detected. HPCD causes the more pronounced stratification in the bilayer, i.e., membrane destabilization near the melting point of CL-rich microphase and, vice versa, a decrease in the lipid mobility in regions with a low CL content. We have studied the effect of HPCD on the interaction of an antibacterial drug, levofloxacin (LV), with the lipid bilayer. It has been found that the complexation of the drug molecules with HPCD leads to an increase in the efficiency of drug adsorption on the surface of the bilayer. This may facilitate the transport of the drug through the bilayer due to the formation of defects in the membrane. LV in the complex with HPCD shows a high antibacterial efficiency in vitro against E. coli, which is not lower as compared with free LV.

Insulin signaling requires glucose to promote lipid anabolism in adipocytes

Adipose tissue is essential for metabolic homeostasis, balancing lipid storage and mobilization based on nutritional status. This is coordinated by insulin, which triggers kinase signaling cascades to modulate numerous metabolic proteins, leading to increased glucose uptake and anabolic processes like lipogenesis. Given recent evidence that glucose is dispensable for adipocyte respiration, we sought to test whether glucose is necessary for insulin-stimulated anabolism. Examining lipogenesis in cultured adipocytes, glucose was essential for insulin to stimulate the synthesis of fatty acids and glyceride-glycerol. Importantly, glucose was dispensable for lipogenesis in the absence of insulin, suggesting that distinct carbon sources are used with or without insulin. Metabolic tracing studies revealed that glucose was required for insulin to stimulate pathways providing carbon substrate, NADPH, and glycerol 3-phosphate for lipid synthesis and storage. Glucose also displaced leucine as a lipogenic substrate and was necessary to suppress fatty acid oxidation. Together, glucose provided substrates and metabolic control for insulin to promote lipogenesis in adipocytes. This contrasted with the suppression of lipolysis by insulin signaling, which occurred independently of glucose. Given previous observations that signal transduction acts primarily before glucose uptake in adipocytes, these data are consistent with a model whereby insulin initially utilizes protein phosphorylation to stimulate lipid anabolism, which is sustained by subsequent glucose metabolism. Consequently, lipid abundance was sensitive to glucose availability, both during adipogenesis and in Drosophila flies in vivo Together, these data highlight the importance of glucose metabolism to support insulin action, providing a complementary regulatory mechanism to signal transduction to stimulate adipose anabolism.

Lipid Droplet Metabolism Across Eukaryotes: Evidence from Yeast to Humans

The lipid droplet (LD) is a highly dynamic organelle that maintains cellular lipid homeostasis in addition to storing energy sources. Current research suggests LDs are responsible for the transportation, storage and lipolysis-driven mobilization of lipids within cells. Here, we review the landscape of evidence for LD involvement in regulating lipid homeostasis. LD interactions with other organelles, particularly the endoplasmic reticulum, mitochondria, lysosomes (or vacuoles in yeast), and peroxisomes, highlight their importance for lipid transfer and metabolism.

Changes in thiamine concentrations, fatty acid composition, and some other lipid-related biochemical indices in Baltic Sea Atlantic salmon (Salmo salar) during the spawning run and pre-spawning fasting

Salmonines in the Baltic Sea and North American lakes suffer from thiamine (vitamin B1) deficiency, which is connected to an abundant lipid-rich diet containing substantial amounts of polyunsaturated fatty acids (PUFAs). In the Baltic region, this is known as the M74 syndrome. It affects both adult salmon (Salmo salar) and especially their offspring, impairing recruitment. However, very little is known about the thiamine and lipid metabolism of salmon during feeding and spawning migrations in the Baltic Sea. In this study, salmon females were sampled along the spawning run from the southern Baltic Proper in four locations at sea and finally at spawning in a river at the Bothnian Bay in a year with insignificant M74 mortality. Changes in concentrations of thiamine and its components in muscle, ovaries, and the liver and other biochemical indices potentially relating to lipid and fatty acid metabolism were investigated. The results provide further evidence of the role of peroxidation of PUFAs in eliciting thiamine deficiency in salmon: During the entire spawning run, the muscle total lipid content decreased by 50%, palmitic acid (16:0) by 62%, and docosahexaenoic acid (DHA, 22:6n-3) by 45%. The concentration of total thiamine decreased significantly until the spawning in the liver and ovaries, 66 and 70% respectively. In the muscle, the proportion of thiamine pyrophosphate of total thiamine increased with the use of muscular lipid stores. There was no trend in the concentration of total carotenoids during the spawning run. The doubling of the concentration of hepatic malondialdehyde indicated peroxidation of PUFAs, and the mobilisation of body lipids suppressed the activity of glucose-6-phosphate dehydrogenase, as consumed dietary lipids would also have done.

Effect of concentration of PEG coated gold nanoparticle on lung surfactant studied with coarse-grained molecular dynamics simulations

The surface modification of nanoparticles can not only change the physical and chemical properties of particles, such as the hydrophilic and hydrophobic properties and surface charges of nanoparticles to a certain extent, but also bring new functions to nanoparticles, such as membrane permeability and targeting. Inhaled nanoparticles (NPs) are experienced by the first biological barrier inside the alveolus known as lung surfactant (LS), consisting of phospholipids and proteins in the form of the monolayer at the air-water interface. Inhaled NPs can reach deep into the lungs and interfere with the biophysical properties of the lung components. The interaction mechanisms of bare gold nanoparticles (AuNPs) with the LS monolayer are not well understood. Coarse-grained molecular dynamics simulations were carried out to have a study on the interactions of PEG coated AuNPs with LS monolayers. It was observed that the interactions of AuNPs and LS components make the monolayer structure deform and change the biophysical properties of LS monolayer. The results also indicate that AuNPs with high concentrations hinder the lowering of the LS surface tension and reduce lateral mobility of lipids. Overall, the simulation results can provide guidance for the design of ligand protected NPs as drug carriers and can identify the nanoparticles potential side effect on lung surfactant.

Functional Omics Identifies Serine Hydrolases That Mobilize Storage Lipids during Rice Seed Germination.

Elucidating proteolipidome dynamics is crucial for understanding the roles of these molecules in plant physiology and disease. Sequence-based functional annotation of the protein is inadequate, since protein activities depend on posttranslational modification. In this study, we applied a gel-free activity-based protein profiling approach to unravel the active lipases, including other Serine hydrolases (SHs), expressed during seed germination in rice (Oryza sativa). We successfully mapped the active sites of 43 active SHs encompassing lipases/esterases, GDSL lipases, proteases, Ser carboxypeptidases, ABHD protein, pectin acetylesterase, and other SHs. The mRNA expression levels of those genes encoding the identified SHs were monitored using microarray analysis. The lipidome analysis revealed distinct patterns of molecular species distribution in individual lipid classes and displayed the metabolic connections between lipid mobilization and rice seedling growth. Changes in the mobilization of storage lipids and their molecular species remodeling were correlated with the expression of the identified lipases and their lipase activity in a time-dependent manner. The physiological significance of the identified SHs was explored during biotic stress with Fusarium verticillioides infection. The fungal infection significantly reduced lipase activity and lipid mobilization, thus impairing the rice seedling. Collectively, our data demonstrate application of the functional proteome strategy along with the shotgun lipidome approach for the identification of active SHs, and thus for deciphering the role of lipid homeostasis during rice seed germination.

Molecular Dynamics Simulations of Cholesterol Effects on the Interaction of hIAPP with Lipid Bilayer.

Fundamental understanding of specific interactions of human islet amyloid polypeptide (hIAPP) with cell membrane is critical for elucidating the underlying pathogenesis of type II diabetes mellitus (T2DM). Membrane cholesterol is known to regulate membrane functions and properties, but its exact role in driving hIAPP-membrane interactions still remains controversial. In this work, we computationally investigated the concentration effect of cholesterol on the adsorption, orientation, and surface interaction of hIAPP oligomers on POPC bilayers containing different amounts of cholesterol (χ = 0, 20, and 40 mol %). Collective MD simulations consistently showed that an increased cholesterol level modulated the structure and dynamics of POPC bilayer, leading to an increase of bilayer thickness, lipid packing order, and surface hydrophobicity but a decrease of lipid mobility. Cholesterol-induced bilayer changes further caused hIAPP oligomer to more preferentially bind to POPC bilayer in the presence of cholesterol via C-terminal residues, in contrast to weak or no binding of hIAPP oligomer on pure POPC bilayers. The cholesterol-enhanced hIAPP-membrane binding is mainly contributed by electrostatic interactions between C-terminal residues and lipid head groups, which may explain the rapid adsorption and aggregation of hIAPP in the presence of cholesterol in cell membranes. This computational work provides some insights into drug development and therapeutic strategies for T2DM by considering cholesterol effects.

Key Process and Factors Controlling the Direct Translocation of Cell-Penetrating Peptide through Bio-Membrane.

Cell-penetrating peptide (CPP) can directly penetrate the cytosol (cytolysis) and is expected to be a potent vector for a drug delivery system (DDS). Although there is general agreement that CPP cytolysis is related to dynamic membrane deformation, a distinctive process has yet to be established. Here, we report the key process and factors controlling CPP cytolysis. To elucidate the task, we have introduced trypsin digestion of adsorbed CPP onto giant unilamellar vesicle (GUV) to quantify the adsorption and internalization (cytolysis) separately. Also, the time-course analysis was introduced for the geometric calculation of adsorption and internalization amount per lipid molecule consisting of GUV. As a result, we found that adsorption and internalization assumed to occur successively by CPP molecule come into contact with membrane lipid. Adsorption is quick to saturate within 10 min, while cytolysis of each CPP on the membrane follows successively. After adsorption is saturated, cytolysis proceeds further linearly by time with a different rate constant that is dependent on the osmotic pressure. We also found that temperature and lipid composition influence cytolysis by modulating lipid mobility. The electrolyte in the outer media is also affected as a chemical mediator to control CPP cytolysis by following the Hoffmeister effect for membrane hydration. These results confirmed the mechanism of cytolysis as temporal and local phase transfer of membrane lipid from Lα to Mesh1, which has punctured bilayer morphologies.