LIVIVO - Search results -

Search results

Result 1 - 10 of total 92

Search options

Article ; Online: Common Mechanism of Activated Catalysis in P-loop Fold Nucleoside Triphosphatases-United in Diversity.

Kozlova, Maria I / Shalaeva, Daria N / Dibrova, Daria V / Mulkidjanian, Armen Y

2022 Volume 12, Issue 10

Abstract: To clarify the obscure hydrolysis mechanism of ubiquitous P-loop-fold nucleoside triphosphatases (Walker NTPases), we analysed the structures of 3136 catalytic sites with bound Mg-NTP complexes or their analogues. Our results are presented in two ... ...

Abstract	To clarify the obscure hydrolysis mechanism of ubiquitous P-loop-fold nucleoside triphosphatases (Walker NTPases), we analysed the structures of 3136 catalytic sites with bound Mg-NTP complexes or their analogues. Our results are presented in two articles; here, in the second of them, we elucidated whether the Walker A and Walker B sequence motifs-common to all P-loop NTPases-could be directly involved in catalysis. We found that the hydrogen bonds (H-bonds) between the strictly conserved, Mg-coordinating Ser/Thr of the Walker A motif ([Ser/Thr]<sup>WA</sup>) and aspartate of the Walker B motif (Asp<sup>WB</sup>) are particularly short (even as short as 2.4 ångströms) in the structures with bound transition state (TS) analogues. Given that a short H-bond implies parity in the pKa values of the H-bond partners, we suggest that, in response to the interactions of a P-loop NTPase with its cognate activating partner, a proton relocates from [Ser/Thr]<sup>WA</sup> to Asp<sup>WB</sup>. The resulting anionic [Ser/Thr]<sup>WA</sup> alkoxide withdraws a proton from the catalytic water molecule, and the nascent hydroxyl attacks the gamma phosphate of NTP. When the gamma-phosphate breaks away, the trapped proton at Asp<sup>WB</sup> passes by the Grotthuss relay via [Ser/Thr]<sup>WA</sup> to beta-phosphate and compensates for its developing negative charge that is thought to be responsible for the activation barrier of hydrolysis.
MeSH term(s)	Nucleoside-Triphosphatase/chemistry ; Nucleoside-Triphosphatase/metabolism ; AAA Domain ; Aspartic Acid ; Protons ; Nucleosides ; Catalysis ; Water/metabolism ; AAA Proteins/metabolism ; Phosphates/metabolism
Chemical Substances	Nucleoside-Triphosphatase (EC 3.6.1.15) ; Aspartic Acid (30KYC7MIAI) ; Protons ; Nucleosides ; Water (059QF0KO0R) ; AAA Proteins (EC 3.6.4.-) ; Phosphates
Language	English
Publishing date	2022-09-22
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2701262-1
ISSN	2218-273X ; 2218-273X
ISSN (online)	2218-273X
ISSN	2218-273X
DOI	10.3390/biom12101346
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

This service is chargeable due to the Delivery terms set by subito. Orders including an article and supplementary material will be classified as separate orders. In these cases, fees will be demanded for each order.

Article ; Online: Common Patterns of Hydrolysis Initiation in P-loop Fold Nucleoside Triphosphatases.

Kozlova, Maria I / Shalaeva, Daria N / Dibrova, Daria V / Mulkidjanian, Armen Y

Biomolecules

2022 Volume 12, Issue 10

Abstract: The P-loop fold nucleoside triphosphate (NTP) hydrolases (also known as Walker NTPases) function as ATPases, GTPases, and ATP synthases, are often of medical importance, and represent one of the largest and evolutionarily oldest families of enzymes. ... ...

Abstract	The P-loop fold nucleoside triphosphate (NTP) hydrolases (also known as Walker NTPases) function as ATPases, GTPases, and ATP synthases, are often of medical importance, and represent one of the largest and evolutionarily oldest families of enzymes. There is still no consensus on their catalytic mechanism. To clarify this, we performed the first comparative structural analysis of more than 3100 structures of P-loop NTPases that contain bound substrate Mg-NTPs or their analogues. We proceeded on the assumption that structural features common to these P-loop NTPases may be essential for catalysis. Our results are presented in two articles. Here, in the first, we consider the structural elements that stimulate hydrolysis. Upon interaction of P-loop NTPases with their cognate activating partners (RNA/DNA/protein domains), specific stimulatory moieties, usually Arg or Lys residues, are inserted into the catalytic site and initiate the cleavage of gamma phosphate. By analyzing a plethora of structures, we found that the only shared feature was the mechanistic interaction of stimulators with the oxygen atoms of gamma-phosphate group, capable of causing its rotation. One of the oxygen atoms of gamma phosphate coordinates the cofactor Mg ion. The rotation must pull this oxygen atom away from the Mg ion. This rearrangement should affect the properties of the other Mg ligands and may initiate hydrolysis according to the mechanism elaborated in the second article.
MeSH term(s)	Nucleoside-Triphosphatase/chemistry ; Nucleoside-Triphosphatase/metabolism ; Hydrolysis ; AAA Domain ; Nucleosides ; Adenosine Triphosphatases/metabolism ; GTP Phosphohydrolases/metabolism ; Adenosine Triphosphate/metabolism ; DNA ; RNA ; Phosphates/metabolism ; AAA Proteins/metabolism ; Oxygen/metabolism
Chemical Substances	Nucleoside-Triphosphatase (EC 3.6.1.15) ; Nucleosides ; Adenosine Triphosphatases (EC 3.6.1.-) ; GTP Phosphohydrolases (EC 3.6.1.-) ; Adenosine Triphosphate (8L70Q75FXE) ; DNA (9007-49-2) ; RNA (63231-63-0) ; Phosphates ; AAA Proteins (EC 3.6.4.-) ; Oxygen (S88TT14065)
Language	English
Publishing date	2022-09-22
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2701262-1
ISSN	2218-273X ; 2218-273X
ISSN (online)	2218-273X
ISSN	2218-273X
DOI	10.3390/biom12101345
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

Article ; Online: Innate Immunity as an Executor of the Programmed Death of Individual Organisms for the Benefit of the Entire Population.

Chernyak, Boris V / Lyamzaev, Konstantin G / Mulkidjanian, Armen Y

International journal of molecular sciences

2021 Volume 22, Issue 24

Abstract: In humans, over-activation of innate immunity in response to viral or bacterial infections often causes severe illness and death. Furthermore, similar mechanisms related to innate immunity can cause pathogenesis and death in sepsis, massive trauma ( ... ...

Abstract	In humans, over-activation of innate immunity in response to viral or bacterial infections often causes severe illness and death. Furthermore, similar mechanisms related to innate immunity can cause pathogenesis and death in sepsis, massive trauma (including surgery and burns), ischemia/reperfusion, some toxic lesions, and viral infections including COVID-19. Based on the reviewed observations, we suggest that such severe outcomes may be manifestations of a controlled suicidal strategy protecting the entire population from the spread of pathogens and from dangerous pathologies rather than an aberrant hyperstimulation of defense responses. We argue that innate immunity may be involved in the implementation of an altruistic programmed death of an organism aimed at increasing the well-being of the whole community. We discuss possible ways to suppress this atavistic program by interfering with innate immunity and suggest that combating this program should be a major goal of future medicine.
MeSH term(s)	Altruism ; Animals ; Apoptosis/immunology ; COVID-19/immunology ; Cell Death/immunology ; Cytokine Release Syndrome/immunology ; Cytokine Release Syndrome/mortality ; Humans ; Immunity, Innate/immunology ; Inflammasomes/immunology ; Inflammation/immunology ; SARS-CoV-2/immunology ; SARS-CoV-2/pathogenicity ; Signal Transduction/immunology
Chemical Substances	Inflammasomes
Language	English
Publishing date	2021-12-15
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms222413480
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

Article ; Online: The microtubule skeleton and the evolution of neuronal complexity in vertebrates.

Trushina, Nataliya I / Mulkidjanian, Armen Y / Brandt, Roland

Biological chemistry

2019 Volume 400, Issue 9, Page(s) 1163–1179

Abstract: The evolution of a highly developed nervous system is mirrored by the ability of individual neurons to develop increased morphological complexity. As microtubules (MTs) are crucially involved in neuronal development, we tested the hypothesis that the ... ...

Abstract	The evolution of a highly developed nervous system is mirrored by the ability of individual neurons to develop increased morphological complexity. As microtubules (MTs) are crucially involved in neuronal development, we tested the hypothesis that the evolution of complexity is driven by an increasing capacity of the MT system for regulated molecular interactions as it may be implemented by a higher number of molecular players and a greater ability of the individual molecules to interact. We performed bioinformatics analysis on different classes of components of the vertebrate neuronal MT cytoskeleton. We show that the number of orthologs of tubulin structure proteins, MT-binding proteins and tubulin-sequestering proteins expanded during vertebrate evolution. We observed that protein diversity of MT-binding and tubulin-sequestering proteins increased by alternative splicing. In addition, we found that regions of the MT-binding protein tau and MAP6 displayed a clear increase in disorder extent during evolution. The data provide evidence that vertebrate evolution is paralleled by gene expansions, changes in alternative splicing and evolution of coding sequences of components of the MT system. The results suggest that in particular evolutionary changes in tubulin-structure proteins, MT-binding proteins and tubulin-sequestering proteins were prominent drivers for the development of increased neuronal complexity.
MeSH term(s)	Animals ; Biological Evolution ; Microtubules/metabolism ; Neurons/metabolism ; Vertebrates ; tau Proteins/metabolism
Chemical Substances	tau Proteins
Language	English
Publishing date	2019-05-18
Publishing country	Germany
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1334659-3
ISSN	1437-4315 ; 1431-6730 ; 1432-0355
ISSN (online)	1437-4315
ISSN	1431-6730 ; 1432-0355
DOI	10.1515/hsz-2019-0149
Database	MEDical Literature Analysis and Retrieval System OnLINE

In stock of ZB MED Cologne/Königswinter

Uc I Zs.50: Show issues

Location:
Je nach Verfügbarkeit (siehe Angabe bei Bestand)
bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular
ab Jg. 2022: Lesesaal (EG)

Order via subito

Details ▾
- See ZB MED holdings
- Order with fees

Article: Activated Q-cycle as a common mechanism for cytochrome bc1 and cytochrome b6f complexes.

Mulkidjanian, Armen Y

Biochimica et biophysica acta

2010 Volume 1797, Issue 12, Page(s) 1858–1868

Abstract: Cytochrome bc(1)-complexes of animals and bacteria (hereafter bc(1)), as well as related cytochrome b(6)f complexes of plants and cyanobacteria (hereafter bf) are dimeric quinol:cytochrome c/plastocyanin oxidoreductases capable of translocating protons ... ...

Abstract	Cytochrome bc(1)-complexes of animals and bacteria (hereafter bc(1)), as well as related cytochrome b(6)f complexes of plants and cyanobacteria (hereafter bf) are dimeric quinol:cytochrome c/plastocyanin oxidoreductases capable of translocating protons across energy-converting membranes. The commonly accepted Q-cycle mechanism suggests that these enzymes oxidize two quinol molecules in their catalytic centers P to yield one quinol molecule in another catalytic center N. Earlier, based upon data on flash-induced redox changes of cytochromes b and c(1), voltage generation, and proton transfer in membrane vesicles of Rhodobacter capsulatus, we have put forward a scheme of an "activated Q-cycle" for the bc(1). The scheme suggests that the bc(1) dimers, being "activated" by injection of electrons from the membrane ubiquinol pool via centers N, steadily contain two electrons in their cytochrome b moieties under physiological conditions, most likely, as a bound semiquinone in center N of one monomer and a reduced high-potential heme b in the other monomer. Then the oxidation of each ubiquinol molecule in centers P of an activated bc(1) should result in a complete catalytic cycle leading to the formation of a ubiquinole molecule in the one of enzyme's centers N and to voltage generation. Here it is argued that a similar pre-loading by two electrons can explain the available data on flash-induced reactions in cytochrome b(6)f-complexes of green plants and cyanobacteria.
MeSH term(s)	Bacterial Proteins/chemistry ; Bacterial Proteins/metabolism ; Benzoquinones/chemistry ; Benzoquinones/metabolism ; Binding Sites ; Cytochrome b6f Complex/chemistry ; Cytochrome b6f Complex/metabolism ; Electron Transport ; Electron Transport Complex III/chemistry ; Electron Transport Complex III/metabolism ; Models, Chemical ; Oxidation-Reduction ; Plant Proteins/chemistry ; Plant Proteins/metabolism ; Rhodobacter capsulatus/enzymology
Chemical Substances	Bacterial Proteins ; Benzoquinones ; Plant Proteins ; quinone (3T006GV98U) ; Cytochrome b6f Complex (9035-40-9) ; Electron Transport Complex III (EC 1.10.2.2)
Language	English
Publishing date	2010-12
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	60-7
ISSN	1879-2596 ; 1879-260X ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650 ; 0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
ISSN (online)	1879-2596 ; 1879-260X ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650
ISSN	0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
DOI	10.1016/j.bbabio.2010.07.008
Database	MEDical Literature Analysis and Retrieval System OnLINE

In stock of ZB MED Cologne/Königswinter

Uc I Zs.247: Show issues

Location:
Je nach Verfügbarkeit (siehe Angabe bei Bestand)
bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular
ab Jg. 2022: Lesesaal (EG)

Order via subito

Details ▾
- See ZB MED holdings
- Order with fees

Article ; Online: Six Functions of Respiration: Isn't It Time to Take Control over ROS Production in Mitochondria, and Aging Along with It?

Skulachev, Vladimir P / Vyssokikh, Mikhail Yu / Chernyak, Boris V / Mulkidjanian, Armen Y / Skulachev, Maxim V / Shilovsky, Gregory A / Lyamzaev, Konstantin G / Borisov, Vitaliy B / Severin, Fedor F / Sadovnichii, Victor A

International journal of molecular sciences

2023 Volume 24, Issue 16

Abstract: Cellular respiration is associated with at least six distinct but intertwined biological functions. (1) biosynthesis of ATP from ADP and inorganic phosphate, (2) consumption of respiratory substrates, (3) support of membrane transport, (4) conversion of ... ...

Abstract	Cellular respiration is associated with at least six distinct but intertwined biological functions. (1) biosynthesis of ATP from ADP and inorganic phosphate, (2) consumption of respiratory substrates, (3) support of membrane transport, (4) conversion of respiratory energy to heat, (5) removal of oxygen to prevent oxidative damage, and (6) generation of reactive oxygen species (ROS) as signaling molecules. Here we focus on function #6, which helps the organism control its mitochondria. The ROS bursts typically occur when the mitochondrial membrane potential (MMP) becomes too high, e.g., due to mitochondrial malfunction, leading to cardiolipin (CL) oxidation. Depending on the intensity of CL damage, specific programs for the elimination of damaged mitochondria (mitophagy), whole cells (apoptosis), or organisms (phenoptosis) can be activated. In particular, we consider those mechanisms that suppress ROS generation by enabling ATP synthesis at low MMP levels. We discuss evidence that the mild depolarization mechanism of direct ATP/ADP exchange across mammalian inner and outer mitochondrial membranes weakens with age. We review recent data showing that by protecting CL from oxidation, mitochondria-targeted antioxidants decrease lethality in response to many potentially deadly shock insults. Thus, targeting ROS- and CL-dependent pathways may prevent acute mortality and, hopefully, slow aging.
MeSH term(s)	Animals ; Reactive Oxygen Species ; Respiration ; Mitochondria ; Aging ; Cardiolipins ; Adenosine Triphosphate ; Mammals
Chemical Substances	Reactive Oxygen Species ; Cardiolipins ; Adenosine Triphosphate (8L70Q75FXE)
Language	English
Publishing date	2023-08-08
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms241612540
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

Article: The Evolution of Tau Phosphorylation and Interactions.

Trushina, Nataliya I / Bakota, Lidia / Mulkidjanian, Armen Y / Brandt, Roland

Frontiers in aging neuroscience

2019 Volume 11, Page(s) 256

Abstract: Tau is a neuronal microtubule-associated protein (MAP) that is involved in the regulation of axonal microtubule assembly. However, as a protein with intrinsically disordered regions (IDRs), tau also interacts with many other partners in addition to ... ...

Abstract	Tau is a neuronal microtubule-associated protein (MAP) that is involved in the regulation of axonal microtubule assembly. However, as a protein with intrinsically disordered regions (IDRs), tau also interacts with many other partners in addition to microtubules. Phosphorylation at selected sites modulates tau's various intracellular interactions and regulates the properties of IDRs. In Alzheimer's disease (AD) and other tauopathies, tau exhibits pathologically increased phosphorylation (hyperphosphorylation) at selected sites and aggregates into neurofibrillary tangles (NFTs). By bioinformatics means, we tested the hypothesis that the sequence of tau has changed during the vertebrate evolution in a way that novel interactions developed and also the phosphorylation pattern was affected, which made tau prone to the development of tauopathies. We report that distinct regions of tau show functional specialization in their molecular interactions. We found that tau's amino-terminal region, which is involved in biological processes related to "membrane organization" and "regulation of apoptosis," exhibited a strong evolutionary increase in protein disorder providing the basis for the development of novel interactions. We observed that the predicted phosphorylation sites have changed during evolution in a region-specific manner, and in some cases the overall number of phosphorylation sites increased owing to the formation of clusters of phosphorylatable residues. In contrast, disease-specific hyperphosphorylated sites remained highly conserved. The data indicate that novel, non-microtubule related tau interactions developed during evolution and suggest that the biological processes, which are mediated by these interactions, are of pathological relevance. Furthermore, the data indicate that predicted phosphorylation sites in some regions of tau, including a cluster of phosphorylatable residues in the alternatively spliced exon 2, have changed during evolution. In view of the "antagonistic pleiotropy hypothesis" it may be worth to take disease-associated phosphosites with low evolutionary conservation as relevant biomarkers into consideration.
Language	English
Publishing date	2019-09-18
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2558898-9
ISSN	1663-4365
ISSN	1663-4365
DOI	10.3389/fnagi.2019.00256
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

Article ; Online: On the origin of life in the zinc world: 1. Photosynthesizing, porous edifices built of hydrothermally precipitated zinc sulfide as cradles of life on Earth.

Mulkidjanian, Armen Y

Biology direct

2009 Volume 4, Page(s) 26

Abstract: ... in the accompanying article (A.Y. Mulkidjanian, M.Y. Galperin, Biology Direct 2009, 4:27).: Conclusion ...

Abstract	Background: The complexity of the problem of the origin of life has spawned a large number of possible evolutionary scenarios. Their number, however, can be dramatically reduced by the simultaneous consideration of various bioenergetic, physical, and geological constraints. Results: This work puts forward an evolutionary scenario that satisfies the known constraints by proposing that life on Earth emerged, powered by UV-rich solar radiation, at photosynthetically active porous edifices made of precipitated zinc sulfide (ZnS) similar to those found around modern deep-sea hydrothermal vents. Under the high pressure of the primeval, carbon dioxide-dominated atmosphere ZnS could precipitate at the surface of the first continents, within reach of solar light. It is suggested that the ZnS surfaces (1) used the solar radiation to drive carbon dioxide reduction, yielding the building blocks for the first biopolymers, (2) served as templates for the synthesis of longer biopolymers from simpler building blocks, and (3) prevented the first biopolymers from photo-dissociation, by absorbing from them the excess radiation. In addition, the UV light may have favoured the selective enrichment of photostable, RNA-like polymers. Falsification tests of this hypothesis are described in the accompanying article (A.Y. Mulkidjanian, M.Y. Galperin, Biology Direct 2009, 4:27). Conclusion: The suggested "Zn world" scenario identifies the geological conditions under which photosynthesizing ZnS edifices of hydrothermal origin could emerge and persist on primordial Earth, includes a mechanism of the transient storage and utilization of solar light for the production of diverse organic compounds, and identifies the driving forces and selective factors that could have promoted the transition from the first simple, photostable polymers to more complex living organisms.
MeSH term(s)	Chemical Precipitation ; Earth, Planet ; Electrons ; Evolution, Chemical ; Geology ; Models, Biological ; Nucleotides/metabolism ; Origin of Life ; Photosynthesis ; Porosity ; Proteins/metabolism ; Reproduction ; Sulfides/chemistry ; Temperature ; Thermodynamics ; Water/chemistry ; Zinc/chemistry ; Zinc Compounds/chemistry
Chemical Substances	Nucleotides ; Proteins ; Sulfides ; Zinc Compounds ; Water (059QF0KO0R) ; Zinc (J41CSQ7QDS) ; zinc sulfide (KPS085631O)
Language	English
Publishing date	2009-08-24
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2221028-3
ISSN	1745-6150 ; 1745-6150
ISSN (online)	1745-6150
ISSN	1745-6150
DOI	10.1186/1745-6150-4-26
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

Article ; Online: Evolution of cell division: from shear mechanics to complex molecular machineries.

Koonin, Eugene V / Mulkidjanian, Armen Y

Cell

2013 Volume 152, Issue 5, Page(s) 942–944

Abstract: Cell division depends on sophisticated molecular machinery. However, wall-less forms of bacteria use a much simpler mechanism that mimics spontaneous division of synthetic lipid vesicles. Mercier et al. (2013) show that this "mechanical" division can be ... ...

Abstract	Cell division depends on sophisticated molecular machinery. However, wall-less forms of bacteria use a much simpler mechanism that mimics spontaneous division of synthetic lipid vesicles. Mercier et al. (2013) show that this "mechanical" division can be activated by increased lipid synthesis. Conceivably, the first cells divided via this route.
Language	English
Publishing date	2013-02-28
Publishing country	United States
Document type	Comment ; Journal Article
ZDB-ID	187009-9
ISSN	1097-4172 ; 0092-8674
ISSN (online)	1097-4172
ISSN	0092-8674
DOI	10.1016/j.cell.2013.02.008
Database	MEDical Literature Analysis and Retrieval System OnLINE

In stock of ZB MED Cologne/Königswinter

Zs.A 1167: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MG 58: Show issues

Order via subito

Details ▾
- See ZB MED holdings
- Order with fees

Article ; Online: A Three-Dimensional Model of the Yeast Transmembrane Sensor Wsc1 Obtained by SMA-Based Detergent-Free Purification and Transmission Electron Microscopy.

Voskoboynikova, Natalia / Karlova, Maria / Kurre, Rainer / Mulkidjanian, Armen Y / Shaitan, Konstantin V / Sokolova, Olga S / Steinhoff, Heinz-Jürgen / Heinisch, Jürgen J

Journal of fungi (Basel, Switzerland)

2021 Volume 7, Issue 2

Abstract: The cell wall sensor Wsc1 belongs to a small family of transmembrane proteins, which are crucial to sustain cell integrity in yeast and other fungi. Wsc1 acts as a mechanosensor of the cell wall integrity (CWI) signal transduction pathway which responds ... ...

Abstract	The cell wall sensor Wsc1 belongs to a small family of transmembrane proteins, which are crucial to sustain cell integrity in yeast and other fungi. Wsc1 acts as a mechanosensor of the cell wall integrity (CWI) signal transduction pathway which responds to external stresses. Here we report on the purification of Wsc1 by its trapping in water-soluble polymer-stabilized lipid nanoparticles, obtained with an amphipathic styrene-maleic acid (SMA) copolymer. The latter was employed to transfer tagged sensors from their native yeast membranes into SMA/lipid particles (SMALPs), which allows their purification in a functional state, i.e., avoiding denaturation. The SMALPs composition was characterized by fluorescence correlation spectroscopy, followed by two-dimensional image acquisition from single particle transmission electron microscopy to build a three-dimensional model of the sensor. The latter confirms that Wsc1 consists of a large extracellular domain connected to a smaller intracellular part by a single transmembrane domain, which is embedded within the hydrophobic moiety of the lipid bilayer. The successful extraction of a sensor from the yeast plasma membrane by a detergent-free procedure into a native-like membrane environment provides new prospects for in vitro structural and functional studies of yeast plasma proteins which are likely to be applicable to other fungi, including plant and human pathogens.
Language	English
Publishing date	2021-02-05
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2784229-0
ISSN	2309-608X ; 2309-608X
ISSN (online)	2309-608X
ISSN	2309-608X
DOI	10.3390/jof7020118
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

To top

More links

Kategorien

Order via subito

More links

Kategorien

Order via subito

More links

Kategorien

Order via subito

More links

Kategorien

In stock of ZB MED Cologne/Königswinter

Order via subito

More links

Kategorien

In stock of ZB MED Cologne/Königswinter

Order via subito

More links

Kategorien

Order via subito

More links

Kategorien

Order via subito

More links

Kategorien

Order via subito

More links

Kategorien

In stock of ZB MED Cologne/Königswinter

Order via subito

More links

Kategorien

Order via subito