Measuring the structural impact of mutations on cytochrome P450 21A2, the major steroid 21-hydroxylase related to congenital adrenal hyperplasia.

Congenital adrenal hyperplasia is an inherited autosomal recessive disorder related to deficient cortisol synthesis. The deficiency of steroid 21-hydroxylase (cytochrome P450 21A2), an enzyme involved in cortisol synthesis, is responsible for ∼95% of cases of congenital adrenal hyperplasia. This metabolic disease exhibits three clinical forms: salt-wasting, simple virilizing, and non-classical form, which are divided according to the degree of severity. In the present study, structural and mutational analyses were performed in order to identify the structural impact of mutations on cytochrome P450 21A2 and correlate them with patient clinical severity. The following mutations were selected: arginine-356 to tryptophan (R356W), proline-30 to leucine (P30L), isoleucine-172 to asparagine (I172N), valine-281 to leucine (V281L), and the null mutation glutamine-318 (Q318X). Our computational approach mapped the location of residues on P450 and identified their implications on enzyme electrostatic potential mapping to progesterone and heme binding pockets. Using molecular dynamics simulations, we analyzed the structural stability of ligand binding and protein structure, as well as possible conformational changes at the catalytic pocket that leads to impairment of enzymatic activity. Our study sheds light on the impact structural mutations have over steroid 21-hydroxylase structure-function in the cell.Communicated by Ramaswamy H. Sarma.

Water spirits within the fishers' worldview: implications for fishing management in Northeast Brazil.

BACKGROUND: Historically, aquatic environments are linked to the worldview of many local people, where there is an interconnection between the natural world, the supernatural, and the social organization. In this study, we provided a discussion on how the supernatural beings that inhabit the freshwater systems interact with artisanal fishers and fishing resources in the riverine community of Parnaíba River middle course, in Mid North of Northeast Brazil. We also provided the implications of these interactions on the fisher's behaviors and how the acculturation process (e.g., introduction of new religions) can affect the fishers' worldview. METHOD: The selection of participants was done through intentional sampling. The content qualitative analysis was carried out to interpret the data from semi-structured interviews with 29 artisanal fishers. RESULTS: The mythical representations that inhabit the aquatic environments in the surveyed area were as follows: Mãe d' água, Cabeça de cuia, Muleque d'água, Visage, Piratinga, Sucuiuiu, and Luz e Arco-íris. These beings have distinctions regarding the form and attributions and can be associated with seasonality (temporal markers) and specific habitats (spatial markers). The respect and fear feeling of the mythical beings were striking among the fishers interviewed. For instance, we have record offering practices in order to obtain protection and success during the fisheries. These practices suggest that there may be local conservationist habits in fisheries management. However, the advancement of urbanization and the introduction of new religions that deny the existence of mythical entities are factors that can generate the acculturation process among the fishers. CONCLUSIONS: It is therefore necessary to carry out more studies in the surveyed area in order to evaluate the existence of possible patterns in the relationship between fisher and mythical beings. This information could confirm the role of mythical beings as environmental regulators. Consequently, it could be considered in the conservationist policies of fishery resources, reinforcing the importance of local knowledge and cultural factors for fishing management approaches.

First homology model of Plasmodium falciparum glucose-6-phosphate dehydrogenase: Discovery of selective substrate analog-based inhibitors as novel antimalarial agents.

In Plasmodium falciparum the bifunctional enzyme glucose-6-phosphate dehydrogenase‒6-phosphogluconolactonase (PfG6PD‒6PGL) is involved in the catalysis of the first reaction of the pentose phosphate pathway. Since this enzyme has a key role in parasite development, its unique structure represents a potential target for the discovery of antimalarial drugs. Here we describe the first 3D structural model of the G6PD domain of PfG6PD‒6PGL. Compared to the human enzyme (hG6PD), the 3D model has enabled the identification of a key difference in the substrate-binding site, which involves the replacement of Arg365 in hG6PD by Asp750 in PfG6PD. In a prospective validation of the model, this critical change has been exploited to rationally design a novel family of substrate analog-based inhibitors that can display the necessary selectivity towards PfG6PD. A series of glucose derivatives featuring an α-methoxy group at the anomeric position and different side chains at position 6 bearing distinct basic functionalities has been synthesized, and their PfG6PD and hG6PD inhibitory activities and their toxicity against parasite and mammalian cells have been assessed. Several compounds displayed micromolar affinity (Ki up to 23 µM), favorable selectivity (up to > 26-fold), and low cytotoxicity. Phenotypic assays with P. falciparum cultures revealed high micromolar IC50 values, likely as a result of poor internalization of the compounds in the parasite cell. Overall, these results endorse confidence to the 3D model of PfG6PD, paving the way for the use of target-based drug design approaches in antimalarial drug discovery studies around this promising target.

Design, synthesis and biological evaluation of N-methyl-N-[(1,2,3-triazol-4-yl)alkyl]propargylamines as novel monoamine oxidase B inhibitors.

Different azides and alkynes have been coupled via Cu-catalyzed 1,3-dipolar Huisgen cycloaddition to afford a novel family of N1- and C5-substituted 1,2,3-triazole derivatives that feature the propargylamine group typical of irreversible MAO-B inhibitors at the C4-side chain of the triazole ring. All the synthesized compounds were evaluated against human MAO-A and MAO-B. Structure-activity relationships and molecular modeling were utilized to gain insight into the structural and chemical features that enhance the binding affinity and selectivity between the two enzyme isoforms. Several lead compounds, in terms of potency (submicromolar to low micromolar range), MAO-B selective recognition, and brain permeability, were identified. One of these leads (MAO-B IC50 of 3.54µM, selectivity MAO-A/MAO-B index of 27.7) was further subjected to reversibility and time-dependence inhibition studies, which disclosed a slow and irreversible inhibition of human MAO-B. Overall, the results support the suitability of the 4-triazolylalkyl propargylamine scaffold for exploring the design of multipotent anti-Alzheimer compounds endowed with irreversible MAO-B inhibitory activity.

What is the role of exotic medicinal plants in local medical systems? A study from the perspective of utilitarian redundancy

Medicinal plants are an important aspect of local medical systems. The composition of a medicinal plant collection is influenced by cultural and environmental factors. Additionally, the functionality of a local medical system can be threatened by the replacement of native species with exotic ones, as well as by cultural factors such as the erosion of knowledge. The objectives of this study are: 1) examine the composition of the medicinal plant collection of two rural communities settled in the caatinga (savanna-like vegetation) of the state of Pernambuco (Brazil); 2) observe the role of exotic plants in the local medical systems; and 3) identify the profile of the species utilized according to the Utilitarian Redundancy Model. Similarities were observed between the medicinal floras of the communities studied, emphasizing the importance of the surrounding biome within the possibilities of species selection, although exotic species appear to contribute by increasing the diversity of species considered in the communities to be medicinal. The native species act broadly among the body systems recognized in the two communities, whereas exotic species act in specific body systems, for which there are few associated native species.

Analysis of the structure of calpain-10 and its interaction with the protease inhibitor SNJ-1715.

Calpain-10 (CAPN10) is a cysteine protease that is activated by intracellular calcium (Ca(2+)) and known to be involved in diseases such as cancer, heart attack, and stroke. A role for the CAPN10 gene in diabetes mellitus type II was recently identified. Hyper activation of the enzyme initiates a series of destructive cycles that can cause irreversible damage to cells. The development of inhibitors may be useful as therapeutic agents for a number of calpainopathies. In this paper, we have used the homology modelling technique to determine the 3D structure of calpain-10 from Homo sapiens. The model of calpain-10 obtained by homology modelling suggests that its active site is conserved among family members and the main interactions are similar to those observed for µ-calpain. Structural analysis revealed that there are small differences in the charge distribution and molecular surface of the enzyme. These differences are probably less dependent on calcium for calpain-10 than they are for µ-calpain. In addition, the ion pair Cys(-)/His(+) formation was observed using of Molecular Dynamics (MD) simulations that were based upon hybrid quantum mechanical/molecular mechanical (QM/MM) approaches. Finally, the binding of the SNJ-1715 inhibitor to calpain-10 was investigated in order to further understand the mechanism of inhibition of calpain-10 by this inhibitor at the molecular level.

Protein-ligand interaction study of CpOGA in complex with GlcNAcstatin.

The GlcNAcstatin is a potent inhibitor of O-glycoprotein 2-acetamino-2-deoxy-ß-D-glucopyranosidase, which has been related with type II diabetes and neurodegenerative disorders. Herein, hybrid quantum mechanics/molecular mechanics, molecular dynamics simulations, and potential of mean force were employed to study the interactions established between GlcNAcstatin and a bacterial O-GlcNAcase enzyme from Clostridium perfringens. The results reveal that the imidazole nitrogen atom of GlcNAcstatin has shown a better interaction with the active site of Clostridium perfringens in its protonated form, which is compatible with a substrate-assisted reaction mechanism involving two conserved aspartate residues (297 and 298). Furthermore, the quantum mechanics/molecular mechanics-molecular dynamics simulations appointed a strong interaction between Asp401, Asp298, and Asp297 residues and the GlcNAcstatin inhibitor, which is in accordance with experimental data. Lastly, these results may contribute to understand the molecular mechanism of inhibition of Clostridium perfringens by GlcNAcstatin inhibitor and, consequently, this study might be useful to design new molecules with more interesting inhibitory activity.

Computational analysis of human OGA structure in complex with PUGNAc and NAG-thiazoline derivatives.

The substitution of serine and threonine residues in nucleocytoplasmic proteins with 2-acetamido-2-deoxy-ß-D-glucopyranose (O-GlcNAc) residues is an essential post-translational modification found in many multicellular eukaryotes. O-glycoprotein 2-acetamino-2-deoxy-ß-D-glucopyranosidase (O-GlcNAcase) hydrolyzes O-GlcNAc residues from post-translationally modified serine/threonine residues of nucleocytoplasmic protein. O-GlcNAc has been implicated in several disease states such as cancer, Alzheimer's disease, and type II diabetes. For this paper, a model of the human O-GlcNAcase (hOGA) enzyme based on the X-ray structures of bacterial Clostridium perfringens (CpNagJ) and Bacteroides thetaiotaomicrometer (BtOGA) homologues has been generated through molecular homology modeling. In addition, molecular docking, molecular dynamics (MD) simulations, and Linear Interaction Energy (LIE) were employed to determine the bind for derivatives of two potent inhibitors: O-(2-acetamido-2-deoxy-D-glucopyranosylidene)amino-N-phenylcarbamate (PUGNAc) and 1,2-dideoxy-2'-methyl-R-D-glucopyranoso-[2,1-d]-Δ2'-thiazoline (NAG-thiazoline), with hOGA. The results show that the binding free energy calculations using the Linear Interaction Energy (LIE) are correlated with inhibition constant values. Therefore, the model of the human O-GlcNAcase (hOGA) obtained here may be used as a target for rational design of new inhibitors.

Molecular modeling of T. rangeli, T. brucei gambiense, and T. evansi sialidases in complex with the DANA inhibitor.

Trypanosomal (trans-) sialidases are enzymes that catalyze the transfer of sialic acid residues between host and parasite glycoconjugates. Herein, we have used homology modeling to construct the 3D structures of sialidases from Trypanosoma brucei and Trypanosoma evansi. Hybrid quantum mechanical/molecular mechanical molecular dynamics simulations were used to determine the interaction energy between the 2-deoxy-2,3-didehydro-N-acetylneuraminic acid inhibitor and the three sialidases studied here. Our results suggest that the two constructed enzymes share the same basic fold motive of the Trypanosoma rangeli crystallographic structure. In addition, quantum mechanical/molecular mechanical molecular dynamics simulations show that the 2-deoxy-2,3-didehydro-N-acetylneuraminic acid inhibitor forms a stronger complex with Trypanosoma rangeli than with Trypanosoma brucei and Trypanosoma evansi sialidases. Finally, the interaction energy by residues shows that the arginine triad plays a decisive role to complex 2-deoxy-2,3-didehydro-N-acetylneuraminic acid with the enzyme through hydrogen bonding.

The use of medicinal plants by migrant people: adaptation, maintenance, and replacement.

Given the importance of studying the knowledge, beliefs, and practices of migrant communities to understand the dynamics of plant resource use, we reviewed the scientific literature concerning the use of medicinal plants by migrant populations engaged in international or long-distance migrations. We considered the importance of two processes: (1) adaptation to the new flora of the host country (i.e., substitution and incorporation of plants in the pharmacopoeia) and (2) continued use and acquisition of the original flora from migrants' home countries (i.e., importation, cultivation, and/or continued use of plants that grow in both host and home environments). We suggest that, depending on the specific context and conditions of migration, different processes that determine the use and/or selection of plants as herbal medicines may become predominant.

A new approach to study medicinal plants with tannins and flavonoids contents from the local knowledge.

AIM OF THE STUDY: The present work tested the power of different methodological strategies for identifying plants that could be interesting in terms of their phenolic compounds (especially flavonoids and tannins) by comparing a new index in which priority-determining criteria are based on the free-listing technique as well as on two randomized methods for choosing plants within an ethnodirected based approach. MATERIALS AND METHODS: The present study was undertaken in the rural area of the municipality of Altinho located in the central region of Pernambuco State, northeastern Brazil. The ethnobotanical survey was divided into three different stages. The first stage was a general survey of 101 individuals on the use and knowledge of medicinal plants within the community. During the second stage local specialists were selected on the basis of the quality and quantity of information they offered during the initial phase of the investigation. The third stage consisted of returning to the specialists a final time in order to apply the free-listing technique. We also assumed that a plant could demonstrate anti-inflammatory and healing effects even without the presence of the compounds of interest of this study. RESULTS: There is a strong association between tannin content and the effects popularly attributed to wound-healing and anti-inflammatory plants. No relationships were observed between plants used by the community to treat inflammation or healing with their flavonoid contents. CONCLUSION: Thus, identifying Caatinga medicinal plants known with anti-inflammatory activity and healing capacities is a good criterion for identifying species with high levels of tannins, although these same criteria are not useful for identifying plants with high flavonoid contents.