Pripravci ljekovitih biljaka za sportaše

Together with training and appropriate rest, diet is a very important part of professional athletes’ lives. This article is about plants preparation that can help sportsmen reach better performances and save them from injuries. The text involves a botanical description, chemical composition, effects, mechanism of action and use of: Salvia hispanica, Beta vulgaris subsp. vulgaris var. conditiva, Panax ginseng, Paullinia cupana, Curcuma longa and Aspalathus linearis. Their benefits can be various, and include providing energy, antiinflamatory, antioxidant, antistress effects, as well as rehydration, muscle regeneration and health preservation

Novel frataxin isoforms may contribute to the pathological mechanism of friedreich ataxia

This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.Friedreich ataxia (FRDA) is an inherited neurodegenerative disease caused by frataxin (FXN) deficiency. The nervous system and heart are the most severely affected tissues. However, highly mitochondria-dependent tissues, such as kidney and liver, are not obviously affected, although the abundance of FXN is normally high in these tissues. In this study we have revealed two novel FXN isoforms (II and III), which are specifically expressed in affected cerebellum and heart tissues, respectively, and are functional in vitro and in vivo. Increasing the abundance of the heart-specific isoform III significantly increased the mitochondrial aconitase activity, while over-expression of the cerebellum-specific isoform II protected against oxidative damage of Fe-S cluster-containing aconitase. Further, we observed that the protein level of isoform III decreased in FRDA patient heart, while the mRNA level of isoform II decreased more in FRDA patient cerebellum compared to total FXN mRNA. Our novel findings are highly relevant to understanding the mechanism of tissue-specific pathology in FRDA.This work was supported by the intramural program of the National Institute of Child Health and Human Development, National Institutes of Health, and in part by Friedreich ataxia research association; by the National Nature Science Foundation of China (NSFC) (No. 31071085), by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry, and by State Key Laboratory of Pharmaceutical Biotechnology (No. ZZYJ-SN-201006). Zvonimir Marelja was supported by a grant from the Studienstiftung des Deutschen Volkes and by Deutscher Akademischer Austauschdienst scholarship. Additional support was obtained from the Deutsche Forschungsgemeinschaft Grant SL1171/5-3

Iron Sulfur and Molybdenum Cofactor Enzymes Regulate the Drosophila Life Cycle by Controlling Cell Metabolism

Iron sulfur (Fe-S) clusters and the molybdenum cofactor (Moco) are present at enzyme sites, where the active metal facilitates electron transfer. Such enzyme systems are soluble in the mitochondrial matrix, cytosol and nucleus, or embedded in the inner mitochondrial membrane, but virtually absent from the cell secretory pathway. They are of ancient evolutionary origin supporting respiration, DNA replication, transcription, translation, the biosynthesis of steroids, heme, catabolism of purines, hydroxylation of xenobiotics, and cellular sulfur metabolism. Here, Fe-S cluster and Moco biosynthesis in Drosophila melanogaster is reviewed and the multiple biochemical and physiological functions of known Fe-S and Moco enzymes are described. We show that RNA interference of Mocs3 disrupts Moco biosynthesis and the circadian clock. Fe-S-dependent mitochondrial respiration is discussed in the context of germ line and somatic development, stem cell differentiation and aging. The subcellular compartmentalization of the Fe-S and Moco assembly machinery components and their connections to iron sensing mechanisms and intermediary metabolism are emphasized. A biochemically active Fe-S core complex of heterologously expressed fly Nfs1, Isd11, IscU, and human frataxin is presented. Based on the recent demonstration that copper displaces the Fe-S cluster of yeast and human ferredoxin, an explanation for why high dietary copper leads to cytoplasmic iron deficiency in flies is proposed. Another proposal that exosomes contribute to the transport of xanthine dehydrogenase from peripheral tissues to the eye pigment cells is put forward, where the Vps16a subunit of the HOPS complex may have a specialized role in concentrating this enzyme within pigment granules. Finally, we formulate a hypothesis that (i) mitochondrial superoxide mobilizes iron from the Fe-S clusters in aconitase and succinate dehydrogenase; (ii) increased iron transiently displaces manganese on superoxide dismutase, which may function as a mitochondrial iron sensor since it is inactivated by iron; (iii) with the Krebs cycle thus disrupted, citrate is exported to the cytosol for fatty acid synthesis, while succinyl-CoA and the iron are used for heme biosynthesis; (iv) as iron is used for heme biosynthesis its concentration in the matrix drops allowing for manganese to reactivate superoxide dismutase and Fe-S cluster biosynthesis to reestablish the Krebs cycle

The N‑Terminus of Iron–Sulfur Cluster Assembly Factor ISD11 Is Crucial for Subcellular Targeting and Interaction with l‑Cysteine Desulfurase NFS1

Assembly of iron–sulfur (FeS) clusters is an important process in living cells. The initial sulfur mobilization step for FeS cluster biosynthesis is catalyzed by l-cysteine desulfurase NFS1, a reaction that is localized in mitochondria in humans. In humans, the function of NFS1 depends on the ISD11 protein, which is required to stabilize its structure. The NFS1/ISD11 complex further interacts with scaffold protein ISCU and regulator protein frataxin, thereby forming a quaternary complex for FeS cluster formation. It has been suggested that the role of ISD11 is not restricted to its role in stabilizing the structure of NFS1, because studies of single-amino acid variants of ISD11 additionally demonstrated its importance for the correct assembly of the quaternary complex. In this study, we are focusing on the N-terminal region of ISD11 to determine the role of N-terminal amino acids in the formation of the complex with NFS1 and to reveal the mitochondrial targeting sequence for subcellular localization. Our in vitro studies with the purified proteins and in vivo studies in a cellular system show that the first 10 N-terminal amino acids of ISD11 are indispensable for the activity of NFS1 and especially the conserved “LYR” motif is essential for the role of ISD11 in forming a stable and active complex with NFS1

The L-Cysteine Desulfurase NFS1 Is Localized in the Cytosol where it Provides the Sulfur for Molybdenum Cofactor Biosynthesis in Humans

<div><p>In humans, the L-cysteine desulfurase NFS1 plays a crucial role in the mitochondrial iron-sulfur cluster biosynthesis and in the thiomodification of mitochondrial and cytosolic tRNAs. We have previously demonstrated that purified NFS1 is able to transfer sulfur to the C-terminal domain of MOCS3, a cytosolic protein involved in molybdenum cofactor biosynthesis and tRNA thiolation. However, no direct evidence existed so far for the interaction of NFS1 and MOCS3 in the cytosol of human cells. Here, we present direct data to show the interaction of NFS1 and MOCS3 in the cytosol of human cells using Förster resonance energy transfer and a split-EGFP system. The colocalization of NFS1 and MOCS3 in the cytosol was confirmed by immunodetection of fractionated cells and localization studies using confocal fluorescence microscopy. Purified NFS1 was used to reconstitute the lacking molybdoenzyme activity of the <i>Neurospora crassa nit-1</i> mutant, giving additional evidence that NFS1 is the sulfur donor for Moco biosynthesis in eukaryotes in general.</p> </div

Immunodetection of NFS1 and MOCS3 after subcellular fractionation of HeLa cells.

<p>Total protein extracts (T), cytosol (C), mitochondria (M), and nucleus (N) were prepared separately from 80–90% confluent HeLa cells to avoid cross contaminations of the compartments. Proteins of each cellular fraction were analyzed by immunoblotting using the following antibodies: anti-NFS1 (<i>top panel</i>), anti-γ-actin as cytosolic marker control (<i>second panel</i>), anti-laminB1 as nuclear marker (<i>third panel</i>), anti-ABCB7 as mitochondrial inner membrane marker (<i>fourth panel</i>), anti-MOCS3 as cytosolic marker (<i>fifth panel</i>), and anti-citrate synthase <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060869#pone.0060869-Kispal1" target="_blank">[6]</a> as mitochondrial matrix marker (<i>bottom</i>).</p

Analysis of NFS1 and MOCS3 interactions by using the split-EGFP system in HeLa cells.

<p>Subcellular EGFP assembly of different split-EGFP fusion proteins was analyzed in HeLa cells by confocal fluorescent microscopy. The following fusion proteins were expressed after cotransfection (assembly of EGFP^1–157 and EGFP^158–238 resulted in a <i>green pseudocolor</i>): <i>A</i>, ISD11-EGFP^1–157 and NFS1-EGFP^158–238; <i>B</i>, MOCS3-EGFP^1–157 and NFS1-EGFP^158–238; <i>C</i>, MOCS3-EGFP^1–157 and NFS1Δ1-55-EGFP^158–238; <i>D</i>, MOCS3-EGFP^1–157 and MOCS3-EGFP^158–238; <i>E</i>, NFS1-EGFP^1–157 and NFS1-EGFP^158–238; <i>F</i>, ISD11-EGFP^1–157 and NFS1Δ1-55-EGFP^158–238. Mitochondria of HeLa cells were visualized with MitoTracker® DeepRed (<i>red</i>) or the nuclei were visualized with DAPI stain (<i>magenta</i>). Merged pictures are shown right (either resulting in a <i>yellow</i> or <i>white color</i>). Scale bars, 20 µm; scale bars in the insets, 2 µm.</p

Analysis of protein-protein interactions between MOCS3 and NFS1Δ1-55 by SPR measurements.

a<p>Proteins were immobilized via amine coupling.</p>b<p>Proteins were injected using KINJECT protocol, injecting samples in a concentration range of 0.3–10 µM. Flow cells were regenerated by injection of 20 mM HCl.</p>c<p>K_D mean values with standard deviation were obtained from 3 independent measurements after global fitting procedures for 1∶1 binding for each measurement.</p>d<p>−, binding not calculated.</p

Fluorescent microscopy of EYFP/ECFP fusion proteins expressed in HeLa cells.

<p>EYFP (<i>green pseudocolor</i>) and ECFP-tagged (<i>red pseudocolor</i>) proteins were analyzed in HeLa cells for subcellular localization and co-localization (the “<i>merge</i>” row resulted in a <i>yellow color</i> when colocalization occurred) by fluorescence confocal microscopy. In addition, a line profile plot (<i>right</i>) shows the pixel intensities of EYFP and ECFP along an arrow (distance in µm) presented in the “merge” figure. <i>A,</i> NFS1-EYFP and ISD11-ECFP (mitochondrial localization); <i>B</i>, NFS1-EYFP (cytosolic localization) and ISD11-ECFP (nuclear localization); <i>C</i>, EYFP-NFS1Δ1-55 and ISD11-ECFP; <i>D</i>, <i>E</i> NFS1-EYFP and ECFP-MOCS3; <i>F</i>, EYFP-NFS1Δ1-55 and ECFP-MOCS3. The figures of <i>panel E</i> are a <i>close-up</i> figure of an indicated area (box) in the figures of <i>panel D</i>. Scale bars, 20 µm (except <i>E,</i> which is 2 µm).</p

Fluorescent microscopy of EYFP/ECFP fusion proteins expressed in HeLa cells.

<p>EYFP (<i>green pseudocolor</i>) and ECFP-tagged (<i>red pseudocolor</i>) proteins were analyzed in HeLa cells for subcellular localization and co-localization (the “<i>merge</i>” row resulted in a <i>yellow color</i> when colocalization occured) by fluorescence confocal microscopy. In addition, a line profile plot (<i>right</i>) shows the pixel intensities of EYFP and ECFP along an arrow (distance in µm) presented in the “merge” figure. <i>A,</i> EYFP-MOCS2A and ECFP-MOCS3; <i>B</i>, EYFP-MOCS2A and NFS1-ECFP; <i>C</i>, EYFP-MOCS2A and ECFP-NFS1Δ1-55; <i>D</i>, EYFP and ECFP. Scale bars, 20 µm.</p