Search results for "102"

2019

Integral membrane proteins of the aquaporin family facilitate rapid water flux across cellular membranes in all domains of life. Although the water-conducting pore is clearly defined in an aquaporin monomer, all aquaporins assemble into stable tetramers. In order to investigate the role of protomer–protomer interactions, we analyzed the activity of heterotetramers containing increasing fractions of mutated monomers, which have an impaired oligomerization propensity and activity. In order to enforce interaction between the protomers, we designed and analyzed a genetically fused homotetramer of GlpF, the aquaglyceroporin of the bacterium Escherichia coli (E. coli). However, increasing fractio…

Spectrophotometric study of DNA interactions with ftorafur and its elementoorganic derivatives

2017

ABSTRACTFtorafur is an antimetabolite antitumour drug successfully used for treatment of various tumours. It is generally accepted that ftorafur is converted to 5-fluoruracil. However, some data indicate direct interactions of the compound with DNA. To test this hypothesis we have performed spectrophotometric study of DNA interactions of ftorafur and some of its elementoorganic derivatives with DNA. UV-VIS spectra of the tested compounds were recorded in absence and presence of increasing amounts of DNA. DNA caused a hypochromic effect in spectra of ftorafur, similar, but weaker effect was observed in 5-fluoruracil spectra. Trimethylgermyl derivative of ftorafur manifested a higher DNA-bind…

Assembly of Transmembrane b-Type Cytochromes and Cytochrome Complexes

2016

Cytochromes are involved in charge-transfer reactions, and many cytochromes contain a transmembrane domain and are part of membrane-localized electron transfer chains. Protoporphyrin IX (heme b) is the first heme product in the tetrapyrrole/heme biosynthesis pathway. In contrast to c-type cytochromes, there is no need for a specialized machinery catalyzing covalent attachment of the heme molecule to a b-type apo-cytochrome, nor is the cofactor further modified, as in a-, d- and o-type cytochromes. Thus, formation of a holo-cytochrome is relatively simple for b-type cytochromes, and this class of proteins probably represents the most ancient members of transmembrane cytochromes. However, ass…

2018

ABC (ATP binding cassette) transporters, ubiquitous in all kingdoms of life, carry out essential substrate transport reactions across cell membranes. Their transmembrane domains bind and translocate substrates and are connected to a pair of nucleotide binding domains, which bind and hydrolyze ATP to energize import or export of substrates. Over four decades of investigations into ABC transporters have revealed numerous details from atomic-level structural insights to their functional and physiological roles. Despite all these advances, a comprehensive understanding of the mechanistic principles of ABC transporter function remains elusive. The human multidrug resistance transporter ABCB1, al…

Echovirus 1 internalization negatively regulates epidermal growth factor receptor downregulation

2016

We have demonstrated previously that the human picornavirus Echovirus 1 (EV1) triggers an infectious internalization pathway that follows closely, but seems to stay separate, from the epidermal growth factor receptor (EGFR) pathway triggered by epidermal growth factor (EGF). Here, we confirmed by using live and confocal microscopy that EGFR and EV1 vesicles are following intimately each other but are distinct entities with different degradation kinetics. We show here that despite being sorted to different pathways and located in distinct endosomes, EV1 inhibits EGFR downregulation. Simultaneous treatment with EV1 and EGF led to an accumulation of EGFR in cytoplasmic endosomes, which was evi…

Environmental aircraft noise aggravates oxidative DNA damage, granulocyte oxidative burst and nitrate resistance inOgg1–/–mice

2020

Background: Large epidemiological studies point towards a link between the incidence of arterial hypertension, ischaemic heart disease, metabolic disease and exposure to traffic noise, supporting t...

Inter- and intraspecific variability in invertebrate acute toxicity response to Arsenic and Fluoride exposure

2018

The adverse effects of arsenic and fluoride exposure on six groups of freshwater invertebrates were investigated. Acute toxicity tests (48-h) with arsenic trioxide (As2NO3) resulted in the following pattern of sensitivity: Daphnia magna 24-h-old = Brachionus patulus 72-h-old = Daphnia. cf. prolata, 21-d-old = D. magna 5-d-old > Heterocypris incongruens juvenile instars > Culex sp. Heterocypris juv. incongruens instars were the second group more tolerant to arsenic and the second group that bioconcentrates arsenic the least. In contrast, invertebrates exposed to sodium fluoride (NaF), showed a different pattern of sensitivity: H. incongruens instars > B. patulus = D. magna 24-h-old > D. cf. …

Cognate HLA absence in trans diminishes human NK cell education

2016

NK cells are innate lymphocytes with protective functions against viral infections and tumor formation. Human NK cells carry inhibitory killer cell Ig-like receptors (KIRs), which recognize distinct HLAs. NK cells with KIRs for self-HLA molecules acquire superior cytotoxicity against HLA– tumor cells during education for improved missing-self recognition. Here, we reconstituted mice with human hematopoietic cells from donors with homozygous KIR ligands or with a mix of hematopoietic cells from these homozygous donors, allowing assessment of the resulting KIR repertoire and NK cell education. We found that co-reconstitution with 2 KIR ligand–mismatched compartments did not alter the frequenc…

NMR Investigation of Structures of G-Protein Coupled Receptor Folding Intermediates

2016

Folding of G-protein coupled receptors (GPCRs) according to the two-stage model (Popot, J. L., and Engelman, D. M. (1990) Biochemistry 29, 4031-4037) is postulated to proceed in 2 steps: partitioning of the polypeptide into the membrane followed by diffusion until native contacts are formed. Herein we investigate conformational preferences of fragments of the yeast Ste2p receptor using NMR. Constructs comprising the first, the first two, and the first three transmembrane (TM) segments, as well as a construct comprising TM1-TM2 covalently linked to TM7 were examined. We observed that the isolated TM1 does not form a stable helix nor does it integrate well into the micelle. TM1 is significant…

Next‐Generation Sequencing‐Based RiboMethSeq Protocol for Analysis of tRNA 2′‐O‐Methylation

2017

Analysis of RNA modifications by traditional physico‐chemical approaches is labor  intensive,  requires  substantial  amounts  of  input  material  and  only  allows  site‐by‐site  measurements.  The  recent  development  of  qualitative  and  quantitative  approaches  based  on   next‐generation sequencing (NGS) opens new perspectives for the analysis of various cellular RNA  species.  The  Illumina  sequencing‐based  RiboMethSeq  protocol  was  initially  developed  and  successfully applied for mapping of ribosomal RNA (rRNA) 2′‐O‐methylations. This method also  gives excellent results in the quantitative analysis of rRNA modifications in different species and  under varying growth condi…