Antibodies against the four isoforms of human plasma membrane Ca2+-ATPase (PMCA) were raised using an N-terminal sequence pump as epitope. The antibodies PMCA 1, 2, and 3 not species-specific, e.g. they also recognized corresponding proteins in rat, whereas that isoform 4 failed to do so.The tissue distribution was estimated by Western blot analysis. Two, PMCA1 PMCA4, expressed all tissues tested (with exception choroid plexus, where former detected). In most signal from protein exceeded...

Tie2 is an endothelium-specific receptor tyrosine kinase that required for both normal embryonic vascular development and tumor angiogenesis thought to play a role in maintenance. However, the signaling pathways responsible function of remain unknown. In this report, we demonstrate p85 subunit phosphatidylinositol 3-kinase (PI3-kinase) associates with association confers functional lipid activity. Mutation 1101 abrogated vitro vivo yeast. was found activate PI3-kinase as demonstrated by...

Several receptor-mediated signal transduction pathways, including EGF and IgE receptor have been proposed to be spatially restricted plasma membrane microdomains. However, the experimental evidence for signaling events in these microdomains is largely based on biochemical fractionation immunocytochemical studies only little known about their spatial dynamics living cells. Here we constructed green fluorescent protein–tagged SH2 domains investigate where when (FcεRI)–mediated tyrosine...

The alternative splicing options and the quantitative tissue distribution of transcripts four currently known human plasma membrane calcium pump (PMCA) genes have been analyzed in seven tissues (cerebral cortex, skeletal heart muscle, stomach, liver, lung, kidney) by polymerase chain reaction on reverse transcribed mRNA with glyceraldehyde-3-phosphate dehydrogenase as internal standard.The mRNAs 1 4 were found to be present similar amounts all tissues, whereas 2 3 expressed a tissue-specific...

The functional significance of the isoform diversity calmodulin-dependent plasma membrane Ca2+-ATPase (PMGA) is largely unknown. To determine whether mRNA synthesis different isoforms enzyme regulated in a differentiation-specific manner, we investigated expression isoform-specific mRNAs muscle and neuronal cells during differentiation by reverse transcription PCR. In rat, ubiquitous PMCA splicing variants 1b 4b formed typical pattern L6 myoblasts, heart-derived cell line H9c2(2-1), two...

The complete structure of the gene for human plasma membrane calcium ATPase isoform 1 (hPMCA1) has been elucidated. protein is encoded by 21 exons present on overlapping clones covering more than 100 kilobases (kb) DNA. An intron over 35 kb separates 5'-untranslated exon from containing translational start codon. entire putative promoter and 5'-flanking region embedded in a CpG island characterized presence numerous Sp1 factor-binding sequences absence TATA box. In accordance with ubiquitous...

While Lyn tyrosine kinase has been shown to be necessary for IgE-receptor (FcepsilonRI)-mediated mast cell activation, the mechanism of activation is not yet understood. Using a micro-electroporation technique quantitatively introduce peptides into cytosol tumor cells, we show that proline-rich preferentially bind Src family SH3 domains block receptor-induced repetitive calcium spikes in concentration dependent manner. The member was likely target, since series phage displaying derived with...

In Switzerland intergovernmental fiscal relations have evolved gradually over 150 years. Like other institutions they reflect first of all the comparatively successful attempt to hold together a society fragmented with respect linguistic and religious diversity along territorial lines. This system is marked by strong decentralization both taxing powers expenditure assignments low equalizing incidence in general combined important redistribution cases high symbolic value. However, internal...

BackgroundDespite the increasing prevalence of postbariatric hypoglycemia (PBH), a late metabolic complication bariatric surgery, our understanding its diverse manifestations remains incomplete.ObjectivesTo contrast parameters glucose-insulin homeostasis in 2 distinct phenotypes PBH (mild versus moderate hypoglycemia) based on nadir plasma glucose.SettingUniversity Hospital (Bern, Switzerland).MethodsTwenty-five subjects with following gastric bypass surgery (age, 41 ± 12 years; body mass...

Abstract Purpose Numerous navigation devices for percutaneous, CT-guided interventions exist and are, due to their advantages, increasingly integrated into the clinical workflow. However, effective training methods ensure safe usage are still lacking. This study compares potential of an augmented reality (AR) application with conventional instructions Cube Navigation System (CNS), hypothesizing enhanced AR, leading safer usage. Methods An AR-tablet app was developed train users puncturing...

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Abstract We retrospectively assessed gluco‐regulatory hormones over 10 h (including two meals) of fully automated closed‐loop insulin delivery using faster (FA) versus standard aspart (IAsp) in adults with type 2 diabetes [n = 15, age 59 ± years, body mass index 34.5 9.1 kg/m , glycated haemoglobin 7.7 1.2% (60 13 mmol/mol)]. Plasma concentration human insulin, IAsp, C‐peptide, glucagon, glucagon‐like peptide 1, glucose‐dependent insulinotropic and tyrosine were measured every 15‐30 min....

Abstract We have investigated the translocation of different GFP-tagged protein domains in response to various receptor stimuli. Confocal fluorescent imaging was used study localization domain before, during and after stimuli applied. A minimal cysteine-rich from kinase C shown be a useful tool localized diacylglycerol changes. Furthermore, calcium-induced plasma membrane C2-domain PKC could receptor-mediated changes near calcium concentration. In addition, pleckstrin homology (PH-domains)...

Abstract We have previously shown that the spatial dynamics of GFP-tagged proteins can be investigated by locally enhancing fluorescence GFP using a short UV-laser pulse. 1 since developed photobleaching and photoenhancement methods to study binding parameters protein domains in intact cells. Here we present an analysis procedure which uses either or measurements determine simultaneously (1) lateral diffusion coefficient (2) dissociation time constant defined for plasma membrane associated,...