scholarly journals Soluble Cyanobacterial Carotenoprotein as a Robust Antioxidant Nanocarrier and Delivery Module

Antioxidants ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 869
Author(s):  
Eugene G. Maksimov ◽  
Alexey V. Zamaraev ◽  
Evgenia Yu. Parshina ◽  
Yury B. Slonimskiy ◽  
Tatiana A. Slastnikova ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Protein Complexes ◽  
Neuroblastoma Cell ◽  
Spectroscopic Properties ◽  
Neuroblastoma Cell Line ◽  
Water Soluble ◽  
Modular Systems ◽  
Pigment Protein Complexes ◽  
Anabaena Sp ◽  
Pcc 7120

To counteract oxidative stress, antioxidants including carotenoids are highly promising, yet their exploitation is drastically limited by the poor bioavailability and fast photodestruction, whereas current delivery systems are far from being efficient. Here we demonstrate that the recently discovered nanometer-sized water-soluble carotenoprotein from Anabaena sp. PCC 7120 (termed AnaCTDH) transiently interacts with liposomes to efficiently extract carotenoids via carotenoid-mediated homodimerization, yielding violet–purple protein samples. We characterize the spectroscopic properties of the obtained pigment–protein complexes and the thermodynamics of liposome–protein carotenoid transfer and demonstrate the delivery of carotenoid echinenone from AnaCTDH into liposomes with an efficiency of up to 70 ± 3%. Most importantly, we show efficient carotenoid delivery to membranes of mammalian cells, which provides protection from reactive oxygen species (ROS). Incubation of neuroblastoma cell line Tet21N in the presence of 1 μM AnaCTDH binding echinenone decreased antimycin A ROS production by 25% (p < 0.05). The described carotenoprotein may be considered as part of modular systems for the targeted antioxidant delivery.

scholarly journals Soluble cyanobacterial carotenoprotein as a robust antioxidant nanocarrier and delivery module

2019 ◽  
Author(s):  
Eugene G. Maksimov ◽  
Alexey V. Zamaraev ◽  
Evgenia Yu. Parshina ◽  
Yury B. Slonimskiy ◽  
Tatiana A. Slastnikova ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Targeted Delivery ◽  
Mammalian Cells ◽  
Protein Complexes ◽  
Natural Antioxidants ◽  
Spectroscopic Properties ◽  
Water Soluble ◽  
Modular Systems ◽  
Long Term Storage ◽  
Pigment Protein Complexes

AbstractTo counteract oxidative stress, antioxidants including carotenoids are highly promising, yet their exploitation is drastically limited by the poor bioavailability and fast photodestruction, whereas current delivery systems are far from being efficient. Here we demonstrate that the recently discovered nanometer-sized water-soluble carotenoprotein from Anabaena (termed CTDH) transiently interacts with liposomes to efficiently extract carotenoids via carotenoid-mediated homodimerization, yielding violet-purple protein samples amenable to lyophilization and long-term storage. We characterize spectroscopic properties of the pigment-protein complexes and thermodynamics of liposome-protein carotenoid transfer and demonstrate the highly efficient delivery of echinenone form CTDH into liposomes. Most importantly, we show carotenoid delivery to membranes of mammalian cells, which provides protection from reactive oxygen species. The described carotenoprotein may be considered as part of modular systems for the targeted antioxidant delivery.Significance statementCarotenoids are excellent natural antioxidants but their delivery to vulnerable cells is challenging due to their hydrophobic nature and susceptibility to degradation. Thus, systems securing antioxidant stability and facilitating targeted delivery are of great interest for the design of medical agents. In this work, we have demonstrated that soluble cyanobacterial carotenoprotein can deliver echinenone into membranes of liposomes and mammalian cells with almost 70 % efficiency, which alleviates the induced oxidative stress. Our findings warrant the robustness of the protein-based carotenoid delivery for studies of carotenoid activities and effects on cell models.

scholarly journals Intercellular transmission of a bacterial cytotoxic prion-like protein in mammalian cells

2019 ◽  
Author(s):  
Aida Revilla-García ◽  
Cristina Fernández ◽  
María Moreno-del Álamo ◽  
Vivian de los Ríos ◽  
Ina M. Vorberg ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Protein Quality ◽  
Horizontal Cell ◽  
Neuroblastoma Cell ◽  
Neuroblastoma Cells ◽  
Neuroblastoma Cell Line ◽  
Human Neuroblastoma ◽  
Intracellular Traffic ◽  
First Time

AbstractRepA is a bacterial protein that builds intracellular amyloid oligomers acting as inhibitory complexes of plasmid DNA replication. When carrying a mutation enhancing its amyloidogenesis (A31V), the N-terminal domain (WH1) generates cytosolic amyloid particles that are inheritable within a bacterial lineage. Such amyloids trigger in bacteria a lethal cascade reminiscent to mitochondria impairment in human cells affected by neurodegeneration. To fulfil all the features of a prion-like protein, horizontal (intercellular) transmissibility remains to be demonstrated for RepA-WH1. Since this is experimentally intractable in bacteria, here we transiently expressed in a murine neuroblastoma cell line the soluble, barely cytotoxic RepA-WH1(WT) and assayed its response to co-incubation with in vitro assembled RepA-WH1(A31V) amyloid fibres. In parallel, cells releasing RepA-WH1(A31V) aggregates were co-cultured with human neuroblastoma cells expressing RepA-WH1(WT). Both the assembled fibres and the extracellular RepA-WH1(A31V) aggregates induce, in the cytosol of recipient cells, the formation of cytotoxic amyloid particles. Mass spectrometry analyses of the proteomes of both types of injured cells point to alterations in mitochondria, protein quality triage, signalling and intracellular traffic.Summary blurbThe horizontal, cell-to-cell spread of a bacterial prion-like protein is shown for the first time in mammalian cells. Amyloid cross-aggregation of distinct variants, and their associated toxicities, follow the same trend found in bacteria, underlining the universality of prion biology.

scholarly journals Insight into the Anticancer Activity of Copper(II) 5-Methylenetrimethylammonium-Thiosemicarbazonates and Their Interaction with Organic Cation Transporters

Biomolecules ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1213
Author(s):  
Miljan N. M. Milunović ◽  
Oleg Palamarciuc ◽  
Angela Sirbu ◽  
Sergiu Shova ◽  
Dan Dumitrescu ◽  
...  
Keyword(s):  
Organic Cation ◽  
Rank Order ◽  
Neuroblastoma Cell ◽  
Neuroblastoma Cell Line ◽  
Water Soluble ◽  
Organic Cation Transporters ◽  
X Ray Diffraction ◽  
Membrane Transport Proteins ◽  
Cation Transporters ◽  
First Time

A series of four water-soluble salicylaldehyde thiosemicarbazones with a positively charged trimethylammonium moiety ([H2LR]Cl, R = H, Me, Et, Ph) and four copper(II) complexes [Cu(HLR)Cl]Cl (1–4) were synthesised with the aim to study (i) their antiproliferative activity in cancer cells and, (ii) for the first time for thiosemicarbazones, the interaction with membrane transport proteins, specifically organic cation transporters OCT1–3. The compounds were comprehensively characterised by analytical, spectroscopic and X-ray diffraction methods. The highest cytotoxic effect was observed in the neuroblastoma cell line SH-5YSY after 24 h exposure and follows the rank order: 3 > 2 > 4 > cisplatin > 1 >> [H2LR]Cl. The copper(II) complexes showed marked interaction with OCT1–3, comparable to that of well-known OCT inhibitors (decynium 22, prazosin and corticosterone) in the cell-based radiotracer uptake assays. The work paves the way for the development of more potent and selective anticancer drugs and/or OCT inhibitors.

scholarly journals Insights into Solution Structures of Photosynthetic Protein Complexes from Small-Angle Scattering Methods

Crystals ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 203
Author(s):  
Maksym Golub ◽  
Adrian Kölsch ◽  
Artem Feoktystov ◽  
Athina Zouni ◽  
Jörg Pieper
Keyword(s):  
Aqueous Solution ◽  
High Resolution ◽  
Photosystem I ◽  
Small Angle ◽  
Protein Complexes ◽  
Photosynthetic Pigment ◽  
Water Soluble ◽  
Model Systems ◽  
Radius Of Gyration ◽  
Pigment Protein Complexes

High-resolution structures of photosynthetic pigment–protein complexes are often determined using crystallography or cryo-electron microscopy (cryo-EM), which are restricted to the use of protein crystals or to low temperatures, respectively. However, functional studies and biotechnological applications of photosystems necessitate the use of proteins isolated in aqueous solution, so that the relevance of high-resolution structures has to be independently verified. In this regard, small-angle neutron and X-ray scattering (SANS and SAXS, respectively) can serve as the missing link because of their capability to provide structural information for proteins in aqueous solution at physiological temperatures. In the present review, we discuss the principles and prototypical applications of SANS and SAXS using the photosynthetic pigment–protein complexes phycocyanin (PC) and Photosystem I (PSI) as model systems for a water-soluble and for a membrane protein, respectively. For example, the solution structure of PSI was studied using SAXS and SANS with contrast matching. A Guinier analysis reveals that PSI in solution is virtually free of aggregation and characterized by a radius of gyration of about 75 Å. The latter value is about 10% larger than expected from the crystal structure. This is corroborated by an ab initio structure reconstitution, which also shows a slight expansion of Photosystem I in buffer solution at room temperature. In part, this may be due to conformational states accessible by thermally activated protein dynamics in solution at physiological temperatures. The size of the detergent belt is derived by comparison with SANS measurements without detergent match, revealing a monolayer of detergent molecules under proper solubilization conditions.

scholarly journals ИССЛЕДОВАНИЕ ФОТОХИМИЧЕСКИХ СВОЙСТВ И ФИЛОГЕНИИ РАСТИТЕЛЬНЫХ БЕЛКОВ СЕМЕЙСТВА WSCP

2020 ◽  
Vol 3 (11(80)) ◽  
pp. 38-43
Author(s):  
Yu. Obuhov ◽  
K. Neverov ◽  
Yu. Maleeva ◽  
M. Krickij
Keyword(s):  
Search Algorithm ◽  
Protein Complexes ◽  
Red Light ◽  
Bioinformatic Analysis ◽  
Class Ii ◽  
Photochemical Activity ◽  
Water Soluble ◽  
Pigment Protein Complexes ◽  
Chlorophyll Protein Complexes ◽  
The Difference

Here we show the ability of the chlorophyll associated with proteins of the WSCP family (Water-Soluble Chlorophyll-binding Proteins) to photosensitize oxidative redox reactions. Irradiation with red light of the recombinant chlorophyll-protein complexes WSCP subclasses IIa and IIb in the presence of an electron donor (NADH) led to oxidation of the donor, i.e., these pigment-protein complexes showed photochemical activity. Meanwhile there was no photodestruction of chlorophyll associated with WSCP, which indicates the photocatalytic nature of the detected redox reaction. The kinetic constants of NADH photooxidation were higher for WSCP subclass IIa (BoWSCP) than for subclass IIb (LvWSCP). To explain the difference in the photosensitizing activity of representatives of different WSCP subclasses, bioinformatic analysis of class II proteins of this family was carried out. For this purpose, we searched for members of the WSCP family in the UniProt protein sequence database using the BLAST search algorithm, followed by their multiple alignment and construction of a phylogenetic tree using the EMBL-EBI Clustal Omega web service and the MEGA7 program. Bioinformatic analysis has confirmed the phylogenetic division of the WSCP class II protein family into two subclasses, previously established on the basis of the difference in their physicochemical properties. It was suggested that phylogeny is related to the photochemical activity of representatives of different subclasses in the WSCP family.

scholarly journals Viologen-phosphorus dendrimers exhibit minor toxicity against a murine neuroblastoma cell line

2013 ◽  
Vol 18 (3) ◽  
Author(s):  
Joanna Lazniewska ◽  
Katarzyna Milowska ◽  
Nadia Katir ◽  
Abdelkim Kadib ◽  
Maria Bryszewska ◽  
...  
Keyword(s):  
Cell Line ◽  
Cellular Response ◽  
Neuroblastoma Cell ◽  
Neuroblastoma Cells ◽  
Neuroblastoma Cell Line ◽  
Water Soluble ◽  
Mitochondrial Activity ◽  
Murine Neuroblastoma ◽  
Biomedical Investigation ◽  
The Impact

AbstractDendrimers containing viologen (derivatives of 4,4′-bipyridyl) units in their structure have been demonstrated to exhibit antiviral activity against human immunodeficiency virus (HIV-1). It has also recently been revealed that novel dendrimers with both viologen units and phosphorus groups in their structure show different antimicrobial, cytotoxic and hemotoxic properties, and have the ability to influence the activity of cholinesterases and to inhibit α-synuclein fibrillation. Since the influence of viologen-phosphorus structures on basic cellular processes had not been investigated, we examined the impact of such macromolecules on the murine neuroblastoma cell line (N2a). We selected three water-soluble viologen-phosphorus (VPD) dendrimers, which differ in their core structure, number of viologen units and number and type of surface groups, and analyzed several aspects of the cellular response. These included cell viability, generation of reactive oxygen species (ROS), alterations in mitochondrial activity, morphological modifications, and the induction of apoptosis and necrosis. The MTT assay results suggest that all of the tested dendrimers are only slightly cytotoxic. Although some changes in ROS formation and mitochondrial function were detected, the three compounds did not induce apoptosis or necrosis. In light of these results, we can assume that the tested VPD are relatively safe for mouse neuroblastoma cells. Although more research on their safety is needed, VPD seem to be promising nanoparticles for further biomedical investigation.

scholarly journals N-Methyl-N-D-fructosyl amphotericin B methyl ester (MF-AME), a novel antifungal agent of low toxicity: monomer/micelle control over selective toxicity.

2000 ◽  
Vol 47 (1) ◽  
pp. 121-131 ◽  
Author(s):  
B Cybulska ◽  
I Gadomska ◽  
J Mazerski ◽  
J G E Borowski ◽  
M Cheron ◽  
...  
Keyword(s):  
Methyl Ester ◽  
Amphotericin B ◽  
Mammalian Cells ◽  
Spectroscopic Properties ◽  
Water Soluble ◽  
Selective Toxicity ◽  
Human Red Blood Cells ◽  
Amphotericin B Methyl Ester

Rational chemical modification of amphotericin B (AMB) led to the synthesis of sterically hindered AMB derivatives. The selected optimal compound, N-methyl-N-D-fructosyl amphotericin B methyl ester (MF-AME) retains the broad spectrum of antifungal activity of the parent antibiotic, and exhibits a two orders of magnitude lower toxicity in vivo and in vitro against mammalian cells. Comparative studies of MF-AME and AMB comprising the determination of the spectroscopic properties of monomeric and self-associated forms of the antibiotics, the investigation of the influence of self-association on toxicity to human red blood cells, and of the antibiotic-sterol interaction were performed. On the basis of the results obtained it can be assumed that the improvement of the selective toxicity of MF-AME could in part be a consequence of the diminished concentration of water soluble oligomers in aqueous medium, and the better ability to differentiate between cholesterol and ergosterol.

Expanding the Natural Products Heterologous Expression Repertoire in the Model Cyanobacterium Anabaena sp. Strain PCC 7120: Production of Pendolmycin and Teleocidin B-4

2019 ◽  
Author(s):  
Patrick Videau ◽  
Kaitlyn Wells ◽  
Arun Singh ◽  
Jessie Eiting ◽  
Philip Proteau ◽  
...  
Keyword(s):  
Natural Products ◽  
Genome Mining ◽  
Gene Clusters ◽  
Combinatorial Biosynthesis ◽  
Test Case ◽  
Biosynthetic Gene ◽  
Biosynthetic Gene Clusters ◽  
Cyanobacterium Anabaena ◽  
Anabaena Sp ◽  
Pcc 7120

Cyanobacteria are prolific producers of natural products and genome mining has shown that many orphan biosynthetic gene clusters can be found in sequenced cyanobacterial genomes. New tools and methodologies are required to investigate these biosynthetic gene clusters and here we present the use of <i>Anabaena </i>sp. strain PCC 7120 as a host for combinatorial biosynthesis of natural products using the indolactam natural products (lyngbyatoxin A, pendolmycin, and teleocidin B-4) as a test case. We were able to successfully produce all three compounds using codon optimized genes from Actinobacteria. We also introduce a new plasmid backbone based on the native <i>Anabaena</i>7120 plasmid pCC7120ζ and show that production of teleocidin B-4 can be accomplished using a two-plasmid system, which can be introduced by co-conjugation.

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