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G-protein coupled receptors (GPCRs) are thought to be proteins with 7-membered transmembrane helical bundles (7TM proteins). Recently, the X-ray structures have been solved for two such proteins, namely for bacteriorhodopsin (BR) and rhodopsin (Rh), the latter being a GPCR. Despite similarities, the structures are different enough to suggest that 3D models for different GPCRs cannot be obtained directly employing 3D structures of BR or Rh as a unique template. The approach to computer modeling of 7TM proteins de­veloped in this work was capable of reproducing the experimental X-ray structure of BR with great accuracy. A combination of helical packing and low-energy conformers for loops most close to the X-ray structure possesses the r.m.s.d. value of 3.13 Ä. Such a level of accuracy for the 3D-structure prediction for a 216-residue protein has not been achieved, so far, by any available ab initio procedure of protein folding. The approach may produce also other energetically consistent combinations of helical bundles and loop con- formers, creating a variety of possible templates for 3D structures of 7TM proteins, in­cluding GPCRs. These templates may provide experimentalists with various plausible op­tions for 3D structure of a given GPCR; in our view, only experiments will determine the fi­nal choice of the most reasonable 3D template.
Ion channels are large transmembrane proteins that are able to conduct small inorganic ions. They are characterized by high selectivity and the ability to gate, i.e. to modify their conductance in response to different stimuli. One of the types of gating follows the ball and chain model, according to which a part of the channel’s protein forms a ball connected with the intracellular side of the channel by a polypeptide chain. The ball is able to modify the conductance of the channel by properly binding to and plugging the channel pore. In this study, the polypeptide ball is treated as a Brownian particle, the movements of which are limited by the length of the chain. The probability density of the ball’s position is resolved by different diffusional operators — parabolic (including the case with drift), hyperbolic, and fractional. We show how those different approaches shed light on different aspects of the movement. We also comment on some features of the survival probabilities (which are ready to be compared with electrophysiological measurements) for issues based on the above operators.
The response of a red blood cell (RBC) to deformation depends on its membrane, a composite of a lipid bilayer and a skeleton, which is a closed, twodimensional network of spectrin tetramers as its bonds. The deformation of the skeleton and its lateral redistribution are studied in terms of the RBC resting state for a fixed geometry of the RBC, partially aspirated into a micropipette. The geometry of the RBC skeleton in its initial state is taken to be either two concentric circles, a references biconcave shape or a sphere. It is assumed that in its initial state the skeleton is distributed laterally in a homogeneous manner with its bonds either unstressed, presenting its stress-free state, or prestressed. The lateral distribution was calculated using a variational calculation. It was assumed that the spectrin tetramer bonds exhibit a linear elasticity. The results showed a significant effect of the initial skeleton geometry on its lateral distribution in the deformed state. The proposed model is used to analyze the measurements of skeleton extension ratios by the method of applying two modes of RBC micropipette aspiration.
 Potassium channels are the most widely distributed class of ion channels. These channels are transmembrane proteins known to play important roles in both normal and pathophysiological functions in all cell types. Various potassium channels are recognised as potential therapeutic targets in the treatment of Parkinson's disease, Alzheimer's disease, brain/spinal cord ischaemia and sepsis. In addition to their importance as therapeutic targets, certain potassium channels are known for their beneficial roles in anaesthesia, cardioprotection and neuroprotection. Some types of potassium channels present in the plasma membrane of various cells have been found in the inner mitochondrial membrane as well. Potassium channels have been proposed to regulate mitochondrial membrane potential, respiration, matrix volume and Ca2+ ion homeostasis. It has been proposed that mitochondrial potassium channels mediate ischaemic preconditioning in various tissues. However, the specificity of a pharmacological agents and the mechanisms underlying their effects on ischaemic preconditioning remain controversial. The following potassium channels from various tissues have been identified in the inner mitochondrial membrane: ATP-regulated (mitoKATP) channel, large conductance Ca2+-regulated (mitoBKCa) channel, intermediate conductance Ca2+-regulated (mitoIKCa) channel, voltage-gated (mitoKv1.3 type) channel, and twin-pore domain (mitoTASK-3) channel. It has been shown that increased potassium flux into brain mitochondria induced by either the mitoKATP channel or mitoBKCa channel affects the beneficial effects on neuronal cell survival under pathological conditions. Recently, differential distribution of mitoBKCa channels has been observed in neuronal mitochondria. These findings may suggest a neuroprotective role for the mitoBKCa channel in specific brain structures. This minireview summarises current data on brain mitochondrial potassium channels and the efforts to identify their molecular correlates.
Tumor necrosis factor-a converting enzyme (TACE) is the first described and best characterized secretase. In this review the structure and the possible roles for TACE are summarized. The substrate specificity and the regulation of TACE activity as well as redundancy and possible cooperations of distinct secretases are also discussed.
Mouse chondrolectin (chodl) was isolated out of the tail tip of fourday old 129/SvJ mice as a by-product of a PCR-based subtractive cDNA library screening. The gene is predominantly expressed in adult skeletal muscle, heart, testes and lungs and in embryonic stadia. Chodl is the mouse homologue of human chondrolectin (CHODL), a gene that encodes for a type Ia transmembrane protein and that is expressed in human testis, prostate, heart and skeletal muscle tissue. CHODL-splice variants (CHODLf, CHODLfΔE, CHODLΔE) are detected in human leukocytes. The proteins of the chondrolectin family belong to the family of C-type lectins. As the members of this protein family are important for a wide array of biological processes, the function of chodl was investigated by searching for its protein interaction partners. The β-subunit of Rab geranylgeranyl transferase (Rabggtb) was isolated 8 times after a complete Sos recruitment system (SRS) screen with the cytoplasmic domain of chodl. The interaction was confirmed with in vitro transcription/translation and co-immunoprecipitation (co-IP) experiments
Molecular cloning and sequencing of a cDNA encoding rabbit presenilin-1 (Ps1) fragment was performed by reverse transcription polymerase chain reaction (RT-PCR) using primers: 5-GGA TGA GCA GCT AAT CTA TAC C-3' and 5-TCC ATT CAG GGA GGT ACT TGA TA-3'. The cDNA fragment revealed 402 nucleotides. The sequence was well conserved and found to be 91, 90, 88, 87 and 78% homologous to that of human, lemur, rat, mouse and chicken, respectively. The cDNA translated into a 130 amino-acid protein fragment. The deduced amino-acid sequence was also well conserved in various species and exhibited 98% similarities with those of rat, lemur and human homologues. However, differences were noticed at residues 145, 168 and 212. This cDNA fragment is quite significant because it is the most conserved portion of Ps1 in various animals and encodes four transmembrane regions (TM2, 3, 4, 5) as defined in human Ps1. Moreover, it includes more than 50% of the sites at which sub­stitutions have been reported in familial Alzheimer's disease (FAD). Therefore, it is suggested that the rabbit can be used as an experimental model for future studies on Ps1 and its physiological functions to work out possible pathways leading to FAD linked neurodegeneration.
Recent studies involving the anion transport protein AE1 (syn. band 3) and glycophorin C in their normal and variant states have demonstrated that these cytoskeletal associated transmembrane proteins play an important role in maintaining the shape and mechanical properties of the human red blood cell. Mutations in the band 3 molecule can lead to a variable outcome on the integrity and physical properties of the erythrocyte. The band 3 Memphis variants and band 3 high transport, for example, appear to have no deleterious effects on the mechanical properties of the red cell whereas total loss of anion transport activity and increased plasma-membrane rigidity is observed with South-east asian ovalocytes. The mechanisms by which mutant band 3 molecules affect the final properties of the cell appear to result from a missasembled membrane domain and an alteration in the protein’s association with the cytoskeleton. Cell shape changes are commonly observed in band 3 mutant cells, manifested as spherocytes, choreoacanthocytes or acanthocytes. Leach phenotype cells, which lack glycophorins C and D and protein p55, when stripped of protein 4.1, have a much reduced affinity for the binding of purified protein 4.1 than normal protein 4.1 depleted cells. Recent studies have shown that protein 4.1 and protein p55 associate directly and on different sites with glycophorin C and that the glycophorin C-protein 4.1-protein p55 complex is required to maintain the shape and mechanical properties of the red cell.
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