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1.
Arch Dis Child ; 2020 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-31969352

RESUMO

OBJECTIVE: Early life adversity has been associated with increased risk of inflammation and inflammation-related diseases in adulthood. This study aimed to examine the association of childhood socioeconomic conditions with chronic low-grade inflammation over adolescence. METHODS: We used information on 2942 members (1507 girls and 1435 boys) of the EPITeen (Epidemiological Health Investigation of Teenagers in Porto) cohort that was established in 2003 in Porto, Portugal, and included 13-year-old adolescents were further evaluated at 17 and 21 years. Mother' and father's education and occupation were used as indicators of childhood socioeconomic conditions. High-sensitivity C reactive protein (hs-CRP) was measured at three points in time (13, 17 and 21 years). hs-CRP levels were categorised in tertiles separately for each wave; chronic low-grade inflammation in adolescence was defined as having hs-CRP levels in the highest tertile in at least two waves and never in the lowest tertile. RESULTS: Prevalence of chronic low-grade inflammation during adolescence was significantly higher among participants with low parental socioeconomic position. Low parental socioeconomic position was associated with chronic low-grade inflammation in adolescence, after adjustment for sex, perinatal and physical environment factors, health-related behaviours and health status in adolescence OR=1.6; 95% CI: 1.1 to 2.4 for lowest versus highest mother's education and OR=1.6; 95% CI: 1.1 to 2.3 for lowest versus highest father's occupation. CONCLUSION: Low childhood socioeconomic conditions are associated with chronic low-grade inflammation during adolescence. Our results suggest that the early life socioeconomic environment has an impact on inflammatory processes over adolescence.

2.
Praxis (Bern 1994) ; 109(1): 23-26, 2020 Jan.
Artigo em Alemão | MEDLINE | ID: mdl-31910756

RESUMO

Socio-Economic Differences in the Lausanne CoLaus Cohort Abstract. The CoLaus study allowed to highlight the existence of broad social inequalities in health among the population of the city of Lausanne. In fact, participants with low socioeconomic status had a higher prevalence of cardio-metabolic risk factors, risk behaviors, sleep disturbances, and higher inflammatory markers compared to the more socio-economically advantaged participants in the study. In most cases, these inequalities are similar to those found in the neighboring cantons and countries.


Assuntos
Doenças Cardiovasculares , Transtornos do Sono-Vigília , Fatores Socioeconômicos , Biomarcadores , Doenças Cardiovasculares/epidemiologia , Estudos de Coortes , Humanos , Fatores de Risco , Transtornos do Sono-Vigília/epidemiologia
3.
Soc Sci Med ; 245: 112685, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31783227

RESUMO

Disadvantaged socioeconomic conditions in childhood heighten systemic inflammatory levels in adulthood; however, life-course mechanisms underlying this association are largely unknown. In the present observational study, we investigated the roles of adulthood socioeconomic and lifestyle factors in mediating this association. Participants were from two prospective Swiss population-based cohorts (N = 5,152, mean age 60 years). We estimated the total effect of paternal occupational position on adult heightened systemic inflammatory levels (C-reactive protein>3 mg/L), and the indirect effects via adulthood socioeconomic positions (SEPs: education and occupational position), financial hardship, and lifestyle factors (body mass index, smoking status, physical inactivity, and alcohol consumption). We estimated odds ratio (OR) and proportion mediated using counterfactual-based mediation models. Individuals whose father had a low occupational position had an OR of 1.51 [95% confidence interval (CI): 1.25, 1.84] for heightened inflammation compared to their more advantaged counterparts. This was jointly mediated (33 [95% CI: 14, 69]%) by adulthood SEPs, whereby the pathway through education followed by occupational position mediated 30 [95% CI: 11, 64]%, while the pathway via occupational position only mediated 3 [95% CI: 4, 13]%. Individuals with the lowest life-course SEPs had an OR of 2.27 [95% CI: 1.71, 2.98] for heightened inflammation compared to having the highest life-course SEPs. This was jointly mediated (63 [95% CI: 44, 97]%) by financial hardship and lifestyle factors. Our study supports a cumulative effect of life-course SEPs on adult heightened systemic inflammation along the pathway paternal occupational position -> education -> adult occupational position. Financial hardship and lifestyle factors in adulthood mediate half of that effect.

4.
Cardiovasc Res ; 2019 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-31754700

RESUMO

AIMS: Sleep disturbances exhibit a strong social patterning, and inadequate sleep has been associated with adverse health outcomes, including cardiovascular disorders (CVD). However, the contribution of sleep to socioeconomic inequalities in CVD is unclear. This study pools data from eight European cohorts to investigate the role of sleep duration in the association between life-course socioeconomic status (SES) and CVD. METHODS AND RESULTS: We used cross-sectional data from eight European cohorts, totalling 111 205 participants. Life-course SES was assessed using father's and adult occupational position. Self-reported sleep duration was categorized into recommended (6-8.5 h/night), long (>8.5 h/night), and short (<6 h/night). We examined two cardiovascular outcomes: coronary heart disease (CHD) and stroke. Main analyses were conducted using pooled data and examined the association between life-course SES and CVD, and the contribution of sleep duration to this gradient using counterfactual mediation. Low father's occupational position was associated with an increased risk of CHD (men: OR = 1.19, 95% CI [1.04; 1.37]; women: OR = 1.25, 95% CI [1.02; 1.54]), with marginal decrease of the gradient after accounting for adult occupational position (men: OR = 1.17, 95% CI [1.02; 1.35]; women: OR = 1.22, 95% CI [0.99; 1.52]), and no mediating effect by short sleep duration. Low adult occupational position was associated with an increased risk of CHD in both men and women (men: OR = 1.48, 95% CI [1.14; 1.92]; women: OR = 1.53, 95% CI [1.04; 2.21]). Short sleep duration meaningfully contributed to the association between adult occupational position and CHD in men, with 13.4% mediation. Stroke did not exhibit a social patterning with any of the variables examined. CONCLUSION: This study suggests that inadequate sleep accounts to a meaningful proportion of the association between adult occupational position and CHD, at least in men. With sleep increasingly being considered an important cardiovascular risk factor in its own terms, our study additionally points to its potential role in social inequalities in cardiovascular disease.

5.
Appl Microbiol Biotechnol ; 103(12): 4801-4812, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30993383

RESUMO

Poly(ethylene terephthalate) (PET) is one of the most widely applied synthetic polymers, but its hydrophobicity is challenging for many industrial applications. Biotechnological modification of PET surface can be achieved by PET hydrolyzing cutinases. In order to increase the adsorption towards their unnatural substrate, the enzymes are fused to carbohydrate-binding modules (CBMs) leading to enhanced activity. In this study, we identified novel PET binding CBMs and characterized the CBM-PET interplay. We developed a semi-quantitative method to detect CBMs bound to PET films. Screening of eight CBMs from diverse families for PET binding revealed one CBM that possesses a high affinity towards PET. Molecular dynamics (MD) simulations of the CBM-PET interface revealed tryptophan residues forming an aromatic triad on the peptide surface. Their interaction with phenyl rings of PET is stabilized by additional hydrogen bonds formed between amino acids close to the aromatic triad. Furthermore, the ratio of hydrophobic to polar contacts at the interface was identified as an important feature determining the strength of PET binding of CBMs. The interaction of CBM tryptophan residues with PET was confirmed experimentally by tryptophan quenching measurements after addition of PET nanoparticles to CBM. Our findings are useful for engineering PET hydrolyzing enzymes and may also find applications in functionalization of PET.


Assuntos
Metabolismo dos Carboidratos , Carboidratos/química , Interações Hidrofóbicas e Hidrofílicas , Polietilenotereftalatos/metabolismo , Triptofano/metabolismo , Sítios de Ligação , Ligações de Hidrogênio , Simulação de Dinâmica Molecular , Ligação Proteica
6.
Methods Mol Biol ; 1880: 17-56, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30610688

RESUMO

Information about the structure and dynamics of proteins is crucial for understanding their physiological functions as well as for the development of strategies to modulate these activities. In this chapter we will describe the work packages required to determine the three-dimensional structures of proteins involved in autophagy by using X-ray crystallography or nuclear magnetic resonance (NMR) spectroscopy. Further we will provide instructions how to perform a molecular dynamics (MD) simulation using GABARAP as example protein.


Assuntos
Proteínas Relacionadas à Autofagia/química , Cristalografia por Raios X/métodos , Ressonância Magnética Nuclear Biomolecular/métodos , Animais , Criopreservação/métodos , Crioprotetores/química , Cristalização/métodos , Humanos , Simulação de Dinâmica Molecular , Conformação Proteica , Software , Proteínas de Transporte Vesicular/química
7.
J Sleep Res ; 28(5): e12799, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30474290

RESUMO

Sleep-disordered breathing is a common condition, related to a higher cardiometabolic and neurocognitive risk. The main risk factors for sleep-disordered breathing include obesity, craniofacial characteristics, male sex and age. However, some studies have suggested that adverse socioeconomic circumstances and lifestyle-related behaviours such as smoking and alcohol use, may also be risk factors for sleep-disordered breathing. Here, we investigate the associations between socioeconomic status and sleep-disordered breathing, as measured by sleep apnea-hypopnea and oxygen desaturation indexes. Furthermore, we assess whether these associations are explained by lifestyle-related factors (smoking, sedentary behaviour, alcohol use and body mass index [BMI]). We used data from the CoLaus|HypnoLaus study, a population-based study including 2162 participants from Lausanne (Switzerland). Socioeconomic status was measured through occupation and education. Sleep-disordered breathing was assessed through polysomnography and measured using the apnea-hypopnea index (AHI: number of apnea/hypopnea events/hr: ≥15/≥30 events), and the ≥3% oxygen desaturation index (ODI: number of oxygen desaturation events/hr: ≥15/≥30 events). Lower occupation and education were associated with higher AHI and ODI (occupation: AHI30, odds ratio (OR) = 1.88, 95% confidence interval (CI) [1.07; 3.31]; ODI30, OR = 2.29, 95% CI [1.19; 4.39]; education: AHI30, OR = 1.21, 95% CI [0.85; 1.72]; ODI30, OR = 1.26, 95% CI [0.83; 1.91]). BMI was associated with socioeconomic status and AHI/ODI, and contributed to the socioeconomic gradient in SDB, with mediation estimates ranging between 43% and 78%. In this Swiss population-based study, we found that low socioeconomic status is a risk factor for sleep-disordered breathing, and that these associations are partly explained by BMI. These findings provide a better understanding of the mechanisms underlying social differences in sleep-disordered breathing and may help implement policies for identifying high-risk profiles for this disorder.

8.
J Am Chem Soc ; 140(46): 15889-15903, 2018 11 21.
Artigo em Inglês | MEDLINE | ID: mdl-30362343

RESUMO

Conformational changes are crucial for the catalytic action of many enzymes. A prototypical and well-studied example is loop opening and closure in triosephosphate isomerase (TIM), which is thought to determine the rate of catalytic turnover in many circumstances. Specifically, TIM loop 6 "grips" the phosphodianion of the substrate and, together with a change in loop 7, sets up the TIM active site for efficient catalysis. Crystal structures of TIM typically show an open or a closed conformation of loop 6, with the tip of the loop moving ∼7 Šbetween conformations. Many studies have interpreted this motion as a two-state, rigid-body transition. Here, we use extensive molecular dynamics simulations, with both conventional and enhanced sampling techniques, to analyze loop motion in apo and substrate-bound TIM in detail, using five crystal structures of the dimeric TIM from Saccharomyces cerevisiae. We find that loop 6 is highly flexible and samples multiple conformational states. Empirical valence bond simulations of the first reaction step show that slight displacements away from the fully closed-loop conformation can be sufficient to abolish most of the catalytic activity; full closure is required for efficient reaction. The conformational change of the loops in TIM is thus not a simple "open and shut" case and is crucial for its catalytic action. Our detailed analysis of loop motion in a highly efficient enzyme highlights the complexity of loop conformational changes and their role in biological catalysis.


Assuntos
Saccharomyces cerevisiae/enzimologia , Triose-Fosfato Isomerase/química , Simulação de Dinâmica Molecular , Estrutura Molecular , Triose-Fosfato Isomerase/metabolismo
9.
Nat Commun ; 9(1): 3900, 2018 09 25.
Artigo em Inglês | MEDLINE | ID: mdl-30254369

RESUMO

Developments in computational chemistry, bioinformatics, and laboratory evolution have facilitated the de novo design and catalytic optimization of enzymes. Besides creating useful catalysts, the generation and iterative improvement of designed enzymes can provide valuable insight into the interplay between the many phenomena that have been suggested to contribute to catalysis. In this work, we follow changes in conformational sampling, electrostatic preorganization, and quantum tunneling along the evolutionary trajectory of a designed Kemp eliminase. We observe that in the Kemp Eliminase KE07, instability of the designed active site leads to the emergence of two additional active site configurations. Evolutionary conformational selection then gradually stabilizes the most efficient configuration, leading to an improved enzyme. This work exemplifies the link between conformational plasticity and evolvability and demonstrates that residues remote from the active sites of enzymes play crucial roles in controlling and shaping the active site for efficient catalysis.


Assuntos
Domínio Catalítico , Projeto Auxiliado por Computador , Evolução Molecular Direcionada , Enzimas/química , Cristalografia por Raios X , Estabilidade Enzimática , Enzimas/genética , Enzimas/metabolismo , Isoxazóis/química , Isoxazóis/metabolismo , Modelos Químicos , Simulação de Dinâmica Molecular , Estrutura Molecular , Eletricidade Estática , Termodinâmica
10.
ACS Catal ; 8(5): 3971-3984, 2018 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-30101036

RESUMO

2-Deoxyribose-5-phosphate aldolase (DERA) catalyzes the reversible conversion of acetaldehyde and glyceraldehyde-3-phosphate into deoxyribose-5-phosphate. DERA is used as a biocatalyst for the synthesis of drugs such as statins and is a promising pharmaceutical target due to its involvement in nucleotide catabolism. Despite previous biochemical studies suggesting the catalytic importance of the C-terminal tyrosine residue found in several bacterial DERAs, the structural and functional basis of its participation in catalysis remains elusive because the electron density for the last eight to nine residues (i.e., the C-terminal tail) is absent in all available crystal structures. Using a combination of NMR spectroscopy and molecular dynamics simulations, we conclusively show that the rarely studied C-terminal tail of E. coli DERA (ecDERA) is intrinsically disordered and exists in equilibrium between open and catalytically relevant closed states, where the C-terminal tyrosine (Y259) enters the active site. Nuclear Overhauser effect distance restraints, obtained due to the presence of a substantial closed state population, were used to derive the solution-state structure of the ecDERA closed state. Real-time NMR hydrogen/deuterium exchange experiments reveal that Y259 is required for efficiency of the proton abstraction step of the catalytic reaction. Phosphate titration experiments show that, in addition to the phosphate-binding residues located near the active site, as observed in the available crystal structures, ecDERA contains previously unknown auxiliary phosphate-binding residues on the C-terminal tail which could facilitate in orienting Y259 in an optimal position for catalysis. Thus, we present significant insights into the structural and mechanistic importance of the ecDERA C-terminal tail and illustrate the role of conformational sampling in enzyme catalysis.

11.
ACS Omega ; 3(4): 3665-3674, 2018 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-30023875

RESUMO

Diamine oxidase (DAO) is an enzyme involved in the regulation of cell proliferation and the immune response. This enzyme performs oxidative deamination in the catabolism of biogenic amines, including, among others, histamine, putrescine, spermidine, and spermine. The mechanistic details underlying the reductive half-reaction of the DAO-catalyzed oxidative deamination which leads to the reduced enzyme cofactor and the aldehyde product are, however, still under debate. The catalytic mechanism was proposed to involve a prototropic shift from the substrate-Schiff base to the product-Schiff base, which includes the rate-limiting cleavage of the Cα-H bond by the conserved catalytic aspartate. Our detailed mechanistic study, performed using a combined quantum chemical cluster approach with empirical valence bond simulations, suggests that the rate-limiting cleavage of the Cα-H bond involves direct hydride transfer to the topaquinone cofactor-a mechanism that does not involve the formation of a Schiff base. Additional investigation of the D373E and D373N variants supported the hypothesis that the conserved catalytic aspartate is indeed essential for the reaction; however, it does not appear to serve as the catalytic base, as previously suggested. Rather, the electrostatic contributions of the most significant residues (including D373), together with the proximity of the Cu2+ cation to the reaction site, lower the activation barrier to drive the chemical reaction.

12.
J R Soc Interface ; 15(144)2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-30021929

RESUMO

Enzymes are dynamic entities, and their dynamic properties are clearly linked to their biological function. It follows that dynamics ought to play an essential role in enzyme evolution. Indeed, a link between conformational diversity and the emergence of new enzyme functionalities has been recognized for many years. However, it is only recently that state-of-the-art computational and experimental approaches are revealing the crucial molecular details of this link. Specifically, evolutionary trajectories leading to functional optimization for a given host environment or to the emergence of a new function typically involve enriching catalytically competent conformations and/or the freezing out of non-competent conformations of an enzyme. In some cases, these evolutionary changes are achieved through distant mutations that shift the protein ensemble towards productive conformations. Multifunctional intermediates in evolutionary trajectories are probably multi-conformational, i.e. able to switch between different overall conformations, each competent for a given function. Conformational diversity can assist the emergence of a completely new active site through a single mutation by facilitating transition-state binding. We propose that this mechanism may have played a role in the emergence of enzymes at the primordial, progenote stage, where it was plausibly promoted by high environmental temperatures and the possibility of additional phenotypic mutations.


Assuntos
Enzimas/genética , Evolução Molecular , Modelos Genéticos , Mutação , Catálise , Domínio Catalítico , Enzimas/metabolismo
13.
Prev Med ; 113: 15-31, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29752959

RESUMO

Unhealthy behaviors and their social patterning have been frequently proposed as factors mediating socioeconomic differences in health. However, a clear quantification of the contribution of health behaviors to the socioeconomic gradient in health is lacking. This study systematically reviews the role of health behaviors in explaining socioeconomic inequalities in health. Published studies were identified by a systematic review of PubMed, Embase and Web-of-Science. Four health behaviors were considered: smoking, alcohol consumption, physical activity and diet. We restricted health outcomes to cardiometabolic disorders and mortality. To allow comparison between studies, the contribution of health behaviors, or the part of the socioeconomic gradient in health that is explained by health behaviors, was recalculated in all studies according to the absolute scale difference method. We identified 114 articles on socioeconomic position, health behaviors and cardiometabolic disorders or mortality from electronic databases and articles reference lists. Lower socioeconomic position was associated with an increased risk of all-cause mortality and cardiometabolic disorders, this gradient was explained by health behaviors to varying degrees (minimum contribution -43%; maximum contribution 261%). Health behaviors explained a larger proportion of the SEP-health gradient in studies conducted in North America and Northern Europe, in studies examining all-cause mortality and cardiovascular disease, among men, in younger individuals, and in longitudinal studies, when compared to other settings. Of the four behaviors examined, smoking contributed the most to social inequalities in health, with a median contribution of 19%. Health behaviors contribute to the socioeconomic gradient in cardiometabolic disease and mortality, but this contribution varies according to population and study characteristics. Nevertheless, our results should encourage the implementation of interventions targeting health behaviors, as they may reduce socioeconomic inequalities in health and increase population health.


Assuntos
Doenças Cardiovasculares/epidemiologia , Comportamentos Relacionados com a Saúde , Disparidades nos Níveis de Saúde , Fumar/epidemiologia , Fatores Socioeconômicos , Doenças Cardiovasculares/mortalidade , Saúde Global , Humanos , Fatores Sexuais
14.
Proteins ; 86(9): 935-944, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29790608

RESUMO

Although the protein backbone is the most fundamental part of the structure, the fine-tuning of side-chain conformations is important for protein function, for example, in protein-protein and protein-ligand interactions, and also in enzyme catalysis. While several benchmarks testing the performance of protein force fields for side chain properties have already been published, they often considered only a few force fields and were not tested against the same experimental observables; hence, they are not directly comparable. In this work, we explore the ability of twelve force fields, which are different flavors of AMBER, CHARMM, OPLS, or GROMOS, to reproduce average rotamer angles and rotamer populations obtained from extensive NMR studies of the 3 J and residual dipolar coupling constants for two small proteins: ubiquitin and GB3. Based on a total of 196 µs sampling time, our results reveal that all force fields identify the correct side chain angles, while the AMBER and CHARMM force fields clearly outperform the OPLS and GROMOS force fields in estimating rotamer populations. The three best force fields for representing the protein side chain dynamics are AMBER 14SB, AMBER 99SB*-ILDN, and CHARMM36. Furthermore, we observe that the side chain ensembles of buried amino acid residues are generally more accurately represented than those of the surface exposed residues.


Assuntos
Antígenos Glicosídicos Associados a Tumores/química , Ubiquitina/química , Simulação de Dinâmica Molecular , Ligação Proteica , Conformação Proteica , Multimerização Proteica , Teoria Quântica
15.
Biophys J ; 114(7): 1614-1623, 2018 04 10.
Artigo em Inglês | MEDLINE | ID: mdl-29642031

RESUMO

Intrinsically disordered proteins dynamically sample a wide conformational space and therefore do not adopt a stable and defined three-dimensional conformation. The structural heterogeneity is related to their proper functioning in physiological processes. Knowledge of the conformational ensemble is crucial for a complete comprehension of this kind of proteins. We here present an approach that utilizes dynamic nuclear polarization-enhanced solid-state NMR spectroscopy of sparsely isotope-labeled proteins in frozen solution to take snapshots of the complete structural ensembles by exploiting the inhomogeneously broadened line-shapes. We investigated the intrinsically disordered protein α-synuclein (α-syn), which plays a key role in the etiology of Parkinson's disease, in three different physiologically relevant states. For the free monomer in frozen solution we could see that the so-called "random coil conformation" consists of α-helical and ß-sheet-like conformations, and that secondary chemical shifts of neighboring amino acids tend to be correlated, indicative of frequent formation of secondary structure elements. Based on these results, we could estimate the number of disordered regions in fibrillar α-syn as well as in α-syn bound to membranes in different protein-to-lipid ratios. Our approach thus provides quantitative information on the propensity to sample transient secondary structures in different functional states. Molecular dynamics simulations rationalize the results.


Assuntos
Espectroscopia de Ressonância Magnética/métodos , alfa-Sinucleína/química , Sequência de Aminoácidos , Simulação de Dinâmica Molecular , Conformação Proteica , Temperatura Ambiente
16.
J Chem Inf Model ; 58(4): 848-858, 2018 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-29522682

RESUMO

Engineering high chemo-, regio-, and stereoselectivity is a prerequisite for enzyme usage in organic synthesis. Cytochromes P450 can oxidize a broad range of substrates, including macrocycles, which are becoming popular scaffolds for therapeutic agents. However, a large conformational space explored by macrocycles not only reduces the selectivity of oxidation but also impairs computational enzyme design strategies based on docking and molecular dynamics (MD) simulations. We present a novel design workflow that uses enhanced-sampling Hamiltonian replica exchange (HREX) MD and focuses on quantifying the substrate binding for suggesting the mutations to be made. This computational approach is applied to P450 BM3 with the aim to shift regioselectively toward one of the numerous possible positions during ß-cembrenediol oxidation. The predictions are experimentally tested and the resulting product distributions validate our design strategy, as single mutations led up to 5-fold regioselectivity increases. We thus conclude that the HREX-MD-based workflow is a promising tool for the identification of positions for mutagenesis aiming at P450 enzymes with improved regioselectivity.


Assuntos
Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Compostos Macrocíclicos/química , Compostos Macrocíclicos/metabolismo , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Engenharia de Proteínas , Domínio Catalítico , Sistema Enzimático do Citocromo P-450/química , Mutagênese , Mutação , Oxirredução , Estereoisomerismo , Especificidade por Substrato , Termodinâmica
17.
Chem Commun (Camb) ; 54(25): 3077-3089, 2018 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-29412205

RESUMO

Phosphate ester hydrolysis is fundamental to many life processes, and has been the topic of substantial experimental and computational research effort. However, even the simplest of phosphate esters can be hydrolyzed through multiple possible pathways that can be difficult to distinguish between, either experimentally, or computationally. Therefore, the mechanisms of both the enzymatic and non-enzymatic reactions have been historically controversial. In the present contribution, we highlight a number of technical issues involved in reliably modeling these computationally challenging reactions, as well as proposing potential solutions. We also showcase examples of our own work in this area, discussing both the non-enzymatic reaction in aqueous solution, as well as insights obtained from the computational modeling of organophosphate hydrolysis and catalytic promiscuity amongst enzymes that catalyze phosphoryl transfer.


Assuntos
Fosfatase Alcalina/metabolismo , Simulação por Computador , Organofosfatos/metabolismo , Fosfatos/metabolismo , Biocatálise , Hidrólise , Modelos Moleculares , Estrutura Molecular , Organofosfatos/química , Fosfatos/química , Teoria Quântica
18.
J Clin Endocrinol Metab ; 103(2): 748-758, 2018 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-29077874

RESUMO

Context: Urinary cadmium (Cd) excretion is associated with cancer and cardiovascular morbidity. A potential mechanism could be disturbance of steroidogenesis in gonads and adrenal glands. Objective: We tested whether urinary excretion of Cd is correlated with that of cortico- and sex steroid metabolites in the general adult population. Setting: The Swiss Kidney Project on Genes in Hypertension is a multicentric, family-based population study. Measures: Urinary excretions of steroid hormone metabolites and Cd were measured with separate day and night collections. Associations were analyzed by mixed linear models. Results: Urinary Cd and testosterone excretions in men were significantly correlated (respective day and night ß values [standard error (SE)], 1.378 [0.242], P < 0.0005; and 1.440 [0.333], P < 0.0005), but not in women [0.333(0.257), P = 0.2; and 0.674 (0.361), P = 0.06]. Urinary Cd and cortisol excretions were positively associated in both sexes [day: ß = 0.475 (SE, 0.157), P = 0.0025, and 0.877 (SE, 0.194), P < 0.0005, respectively; night: ß = 0.875 (SE, 0.253), P < 0.0005 and 1.183 (SE, 0.277), P = 0.00002, respectively]. Cd excretion was correlated with mineralocorticoid metabolites excretion, except tetrahydroaldosterone, in both sexes (P < 0.01). There was an independent effect of Cd on sex hormone and corticosteroid synthesis and an interdependent effect on gluco- and mineralcorticoid production. Conclusion: Our findings provide evidence for a global stimulating effect on steroid synthesis already at low-dose Cd exposure. These findings might explain the association of Cd with diseases such as steroid-sensitive cancers or metabolic disorders.


Assuntos
Corticosteroides/metabolismo , Cádmio/urina , Hormônios Esteroides Gonadais/metabolismo , Hipertensão/metabolismo , Adulto , Idoso , Aldosterona/análogos & derivados , Aldosterona/urina , Estudos de Coortes , Família , Feminino , Hormônios Esteroides Gonadais/urina , Humanos , Hipertensão/urina , Rim/metabolismo , Masculino , Pessoa de Meia-Idade , Mineralocorticoides/urina , Testosterona/urina
19.
J Am Chem Soc ; 139(30): 10514-10525, 2017 08 02.
Artigo em Inglês | MEDLINE | ID: mdl-28683550

RESUMO

Triosephosphate isomerase (TIM) is a proficient catalyst of the reversible isomerization of dihydroxyacetone phosphate (DHAP) to d-glyceraldehyde phosphate (GAP), via general base catalysis by E165. Historically, this enzyme has been an extremely important model system for understanding the fundamentals of biological catalysis. TIM is activated through an energetically demanding conformational change, which helps position the side chains of two key hydrophobic residues (I170 and L230), over the carboxylate side chain of E165. This is critical both for creating a hydrophobic pocket for the catalytic base and for maintaining correct active site architecture. Truncation of these residues to alanine causes significant falloffs in TIM's catalytic activity, but experiments have failed to provide a full description of the action of this clamp in promoting substrate deprotonation. We perform here detailed empirical valence bond calculations of the TIM-catalyzed deprotonation of DHAP and GAP by both wild-type TIM and its I170A, L230A, and I170A/L230A mutants, obtaining exceptional quantitative agreement with experiment. Our calculations provide a linear free energy relationship, with slope 0.8, between the activation barriers and Gibbs free energies for these TIM-catalyzed reactions. We conclude that these clamping side chains minimize the Gibbs free energy for substrate deprotonation, and that the effects on reaction driving force are largely expressed at the transition state for proton transfer. Our combined analysis of previous experimental and current computational results allows us to provide an overview of the breakdown of ground-state and transition state effects in enzyme catalysis in unprecedented detail, providing a molecular description of the operation of a hydrophobic clamp in triosephosphate isomerase.


Assuntos
Fosfato de Di-Hidroxiacetona/metabolismo , Gliceraldeído 3-Fosfato/metabolismo , Simulação de Dinâmica Molecular , Triose-Fosfato Isomerase/metabolismo , Biocatálise , Fosfato de Di-Hidroxiacetona/química , Gliceraldeído 3-Fosfato/química , Interações Hidrofóbicas e Hidrofílicas , Conformação Molecular , Saccharomyces cerevisiae/enzimologia , Termodinâmica , Triose-Fosfato Isomerase/química , Triose-Fosfato Isomerase/genética
20.
Am J Public Health ; 107(4): e1-e12, 2017 04.
Artigo em Inglês | MEDLINE | ID: mdl-28207328

RESUMO

BACKGROUND: A poorer quality diet among individuals with low socioeconomic status (SES) may partly explain the higher burden of noncommunicable disease among disadvantaged populations. Because there is a link between sodium intake and noncommunicable diseases, we systematically reviewed the current evidence on the social patterning of sodium intake. OBJECTIVES: To conduct a systematic review and a meta-analysis of the evidence on the association between SES and sodium intake in healthy adult populations of high-income countries. SEARCH METHODS: We followed the PRISMA-Equity guidelines in conducting a literature search that ended June 3, 2016, via MEDLINE, Embase, and SciELO. We imposed no publication date limits. SELECTION CRITERIA: We considered only peer-reviewed articles meeting the following inclusion criteria: (1) reported a measure of sodium intake disaggregated by at least 1 measure of SES (education, income, occupation, or any other socioeconomic indicator); (2) were written in English, Spanish, Portuguese, French, or Italian; and (3) were conducted in a high-income country as defined by the World Bank (i.e., per capita national gross income was higher than $12 746). We also excluded articles that exclusively sampled low-SES individuals, pregnant women, children, adolescents, elderly participants, or diseased patients or that reported results from a trial or intervention. DATA COLLECTION AND ANALYSIS: As summary measures, we extracted (1) the direction (positive, negative, or neutral) and the magnitude of the association between each SES indicator and sodium intake, and (2) the estimated sodium intake according to SES level. When possible and if previously unreported, we calculated the magnitude of the relative difference in sodium intake between high- and low-SES groups for each article, applying this formula: ([value for high-SES group - value for low-SES group]/[value for high-SES group]) × 100. We considered an association significant if reported as such, and we set an arbitrary 10% relative difference as clinically relevant and significant. We conducted a meta-analysis of the relative difference in sodium intake between high- and low-SES groups. We included articles in the meta-analysis if they reported urine-based sodium estimates and provided the total participant numbers in the low- and high-SES groups, the estimated sodium intake means for each group (in mg/day or convertible units), and the SDs (or transformable measures). We chose a random-effects model to account for both within-study and between-study variance. MAIN RESULTS: Fifty-one articles covering 19 high-income countries met our inclusion criteria. Of these, 22 used urine-based methods to assess sodium intake, and 30 used dietary surveys. These articles assessed 171 associations between SES and sodium intake. Among urine-based estimates, 67% were negative (higher sodium intake in people of low SES), 3% positive, and 30% neutral. Among diet-based estimates, 41% were negative, 21% positive, and 38% neutral. The random-effects model indicated a 14% relative difference between low- and high-SES groups (95% confidence interval [CI] = -18, -9), corresponding to a global 503 milligrams per day (95% CI = 461, 545) of higher sodium intake among people of low SES. CONCLUSIONS: People of low SES consume more sodium than do people of high SES, confirming the current evidence on socioeconomic disparities in diet, which may influence the disproportionate noncommunicable disease burden among disadvantaged socioeconomic groups. Public Health Implications. It is necessary to focus on disadvantaged populations to achieve an equitable reduction in sodium intake to a population mean of 2 grams per day as part of the World Health Organization's target to achieve a 25% relative reduction in noncommunicable disease mortality by 2025.


Assuntos
Países Desenvolvidos , Fatores Socioeconômicos , Sódio na Dieta/administração & dosagem , Adulto , Feminino , Humanos , Masculino
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