Using PCR and inverse PCR techniques we obtained a 4,498 bp nucleotide sequence FN424076 encompassing the complete coding sequence of the porcine insulin receptor substrate 4 (IRS4) gene and its proximal promoter. The 1,269 amino acid porcine protein deduced from the nucleotide sequence shares 92% identity with the human IRS4 and possesses the same domains and the same number of tyrosine phosphorylation motifs as the human protein. We detected substitution FN424076:g.96C

• MeSH
• celogenomová asociační studie MeSH
• DNA primery genetika   MeSH
• fenotyp MeSH
• jednonukleotidový polymorfismus genetika   MeSH
• klonování DNA MeSH
• lineární modely MeSH
• mapování chromozomů MeSH
• molekulární sekvence - údaje MeSH
• polymerázová řetězová reakce MeSH
• prasata genetika   MeSH
• proteiny insulinového receptorového substrátu genetika   MeSH
• sekvence nukleotidů MeSH
• sekvenční analýza DNA MeSH
• sekvenční homologie MeSH
• tělesné váhy a míry MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

To investigate structure-function relationships of cytochromes P450 (CYP), 3-azidiamantane was employed for photoaffinity labeling of rabbit microsomal CYP2B4. Four diamantane labeled tryptic fragments were identified by mass spectrometry and sequencing: peptide I (Leu359-Lys373), peptide II (Leu30-Arg48), peptide III (Phe127-Arg140), and peptide IV (Arg434-Arg443). Their positions were projected into CYP2B4 model structures and compared with substrate binding sites, proposed by docking of diamantane. We identified novel binding regions outside the active site of CYP2B4. One of them, defined with diamantane modified Arg133, marks a possible entrance to the active site from the heme proximal face. In addition to crystal structures of CYP2B4 chimeras and molecular dynamics simulations, our data of photoaffinity labeling of the full CYP2B4 molecule provide further insight into functional and structural aspects of substrate binding.

• MeSH
• adamantan analogy a deriváty   chemie   MeSH
• aromatické hydroxylasy chemie   ultrastruktura   MeSH
• chemické modely MeSH
• financování organizované MeSH
• fluorescenční mikroskopie metody   MeSH
• fotochemie metody   MeSH
• konformace proteinů MeSH
• molekulární modely MeSH
• počítačová simulace MeSH
• substrátová specifita MeSH
• vazba proteinů MeSH
• vazebná místa MeSH

Histone deacetylase 6 (HDAC6) is a multidomain cytosolic hydrolase acting mostly on nonhistone protein substrates. Investigations of the substrate specificity of HDAC6 are confounded by the presence of 2 catalytically active deacetylase domains (DD1 and DD2). In this study, acetylome peptide microarrays and peptide libraries were used to map the substrate specificity of DD1 and DD2 of human HDAC6. The results show that DD1 is solely responsible for the deacetylation of substrates harboring the acetyllysine at their C terminus, whereas DD2 exclusively deacetylates peptides with an internal acetyllysine residue. Also, statistical analysis of the deacetylation data revealed amino acid preferences at individual positions flanking the acetyllysine, where glycine and arginine residues are favored at positions N-terminal to the central acetyllysine; negatively charged glutamate is strongly disfavored throughout the sequence. Finally, the deacylation activity of HDAC6 was profiled by using a panel of acyl derivatives of the optimized peptide substrate and showed that HDAC6 acts as a proficient deformylase. Our data thus offer a detailed insight into the substrate preferences of the individual HDAC6 domains at the peptide level, and these findings can in turn help in elucidating the biologic roles of the enzyme and facilitate the development of new domain-specific inhibitors as research tools or therapeutic agents.-Kutil, Z., Skultetyova, L., Rauh, D., Meleshin, M., Snajdr, I., Novakova, Z., Mikesova, J., Pavlicek, J., Hadzima, M., Baranova, P., Havlinova, B., Majer, P., Schutkowski, M., Barinka, C. The unraveling of substrate specificity of histone deacetylase 6 domains using acetylome peptide microarrays and peptide libraries.

• MeSH
• HEK293 buňky MeSH
• histondeacetylasa 6 chemie   metabolismus   MeSH
• katalytická doména * MeSH
• lidé MeSH
• lysin chemie   metabolismus   MeSH
• peptidové fragmenty chemie   metabolismus   MeSH
• statická elektřina MeSH
• substrátová specifita MeSH
• vazba proteinů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Plant mitogen-activated protein kinases (MAPKs) constitute a network of signaling cascades responsible for transducing extracellular stimuli and decoding them to dedicated cellular and developmental responses that shape the plant body. Over the last decade, we have accumulated information about how MAPK modules control the development of reproductive tissues and gametes and the embryogenic and postembryonic development of vegetative organs such as roots, root nodules, shoots, and leaves. Of key importance to understanding how MAPKs participate in developmental and environmental signaling is the characterization of their subcellular localization, their interactions with upstream signal perception mechanisms, and the means by which they target their substrates. In this review, we summarize the roles of MAPK signaling in the regulation of key plant developmental processes, and we survey what is known about the mechanisms guiding the subcellular compartmentalization of MAPK modules.

• MeSH
• MAP kinasový signální systém MeSH
• mitogenem aktivované proteinkinasy metabolismus   MeSH
• orgánová specificita MeSH
• rostlinné buňky enzymologie   MeSH
• rostliny enzymologie   MeSH
• vývojová biologie * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

BACKGROUND: Patients with coronary artery disease (CAD) may have ventricular tachycardia (VT) from a separate nonischemic process. Catheter ablation in these patients can be misguided by abnormalities of coronary arteries. OBJECTIVE: To identify (1) the prevalence of unanticipated nonischemic VT in patients with known CAD presenting with VT and (2) the substrate and VT characteristics of this unique subset of patients. METHODS: We examined consecutive patients referred for VT catheter ablation who had a history of myocardial infarction and angiography documented CAD with presumed ischemic VT. Patients with low-voltage zones and/or VT origin inconsistent with CAD distribution were included for further analysis. RESULTS: Of 732 patients, 9 (1.2%) (7 men; median age 74 years; ejection fraction 30%) fulfilled inclusion criteria. Endocardial left ventricular scar inconsistent with CAD distribution was found in 8 patients. In 1 patient, only epicardial left ventricular scar was found. The distribution of low voltage (<1.5 mV) was predominantly around the aortic and mitral valves. Thirty-one VTs were induced in 8 patients. Most VTs had right bundle branch block (68%); of these VTs, 67% had an R/S transition zone later than lead V4 consistent with basal VT origin. Epicardial ablation was necessary in 2 patients. During follow-up (30 [25-39] months), 7 of 9 patients (78%) were free of recurrent VT. CONCLUSIONS: A small but important subgroup of patients with CAD and VT has a nonischemic substrate/etiology for VT. The presence of multiple VTs with basal origin suggests a potential nonischemic perivalvular substrate and possible need for epicardial VT ablation.

• MeSH
• kardiomyopatie komplikace   epidemiologie   patofyziologie   MeSH
• katetrizační ablace MeSH
• komorová tachykardie komplikace   epidemiologie   chirurgie   MeSH
• lidé MeSH
• mapování potenciálů tělesného povrchu MeSH
• následné studie MeSH
• nemoci koronárních tepen komplikace   diagnóza   patofyziologie   MeSH
• prevalence MeSH
• prognóza MeSH
• senioři MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

Stress-activated plant mitogen-activated protein (MAP) kinase pathways play roles in growth adaptation to the environment by modulating cell division through cytoskeletal regulation, but the mechanisms are poorly understood. We performed protein interaction and phosphorylation experiments with cytoskeletal proteins, mass spectrometric identification of MPK6 complexes and immunofluorescence analyses of the microtubular cytoskeleton of mitotic cells using wild-type, mpk6-2 mutant and plants overexpressing the MAP kinase-inactivating phosphatase, AP2C3. We showed that MPK6 interacted with γ-tubulin and co-sedimented with plant microtubules polymerized in vitro. It was the active form of MAP kinase that was enriched with microtubules and followed similar dynamics to γ-tubulin, moving from poles to midzone during the anaphase-to-telophase transition. We found a novel substrate for MPK6, the microtubule plus end protein, EB1c. The mpk6-2 mutant was sensitive to 3-nitro-l-tyrosine (NO2 -Tyr) treatment with respect to mitotic abnormalities, and root cells overexpressing AP2C3 showed defects in chromosome segregation and spindle orientation. Our data suggest that the active form of MAP kinase interacts with γ-tubulin on specific subsets of mitotic microtubules during late mitosis. MPK6 phosphorylates EB1c, but not EB1a, and has a role in maintaining regular planes of cell division under stress conditions.

• MeSH
• anafáze účinky léků   MeSH
• aparát dělícího vřeténka účinky léků   metabolismus   MeSH
• Arabidopsis cytologie   účinky léků   enzymologie   MeSH
• butadieny farmakologie   MeSH
• cytokineze účinky léků   MeSH
• extracelulárním signálem regulované MAP kinasy metabolismus   MeSH
• fosforylace účinky léků   MeSH
• fyziologický stres * účinky léků   MeSH
• kinetochory účinky léků   metabolismus   MeSH
• meristém cytologie   účinky léků   metabolismus   MeSH
• mikrotubuly účinky léků   metabolismus   MeSH
• mitogenem aktivované proteinkinasy metabolismus   MeSH
• nitrily farmakologie   MeSH
• nitrosace účinky léků   MeSH
• proliferace buněk účinky léků   MeSH
• proteiny asociované s mikrotubuly metabolismus   MeSH
• proteiny huseníčku metabolismus   MeSH
• rostlinné buňky účinky léků   metabolismus   MeSH
• segregace chromozomů účinky léků   MeSH
• telofáze účinky léků   MeSH
• tubulin metabolismus   MeSH
• tyrosin analogy a deriváty   farmakologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Cdc14 is a member of the dual-specificity phosphatase family, which is essential for faithful cell cycle progression in eukaryotic cells of different origin. The function of human Cdc14A (hCdc14A), however, has not been fully elucidated as only few physiological substrates have been identified. To gain insight into the biological role of Cdc14A, we performed a yeast two-hybrid screen designed to isolate substrates of this human phosphatase. Using this genetic approach, we here report the identification of Erk3, an atypical mitogen-activated protein kinase (MAPK), as a specific binding partner of hCdc14A. GST pull-down assays show that Erk3 interacts directly with hCdc14A in vitro via its unique C-terminal domain. Furthermore, biochemical analysis reveals that hCdc14A can remove cyclin-dependent kinase (Cdk)-mediated phosphorylation of Erk3 in vitro raising the possibility that Erk3 may be a potential substrate for hCdc14A in vivo. Consistent with a physiologically relevant cross-talk in vivo, we find that Cdc14A forms a stable complex with Erk3 in human cells independent of its intrinsic phosphatase activity but mediated by its regulatory C-terminal domain. We show that hCdc14A impacts the emerging signaling pathway between Erk3 and MK5, a MAPK-activated protein kinase. We document that hCdc14A upregulation leads to redistribution of the Erk3 substrate MK5 from the nucleus to the cytoplasm. In addition, we find that hCdc14A stabilizes complex formation between Erk3 and its binding partner cyclin D3, a D-type cyclin implicated in both cellular proliferation and differentiation. Collectively, our findings suggest an intimate functional relationship between the Cdc14A phosphatase and the Erk3 kinase in signaling pathways that regulate key cell-fate decisions in human cells.

• MeSH
• buněčný cyklus fyziologie   MeSH
• fosfatasy * genetika   fyziologie   MeSH
• lidé MeSH
• MAP kinasový signální systém fyziologie   MeSH
• mitogenem aktivovaná proteinkinasa 6 * genetika   fyziologie   MeSH
• nádorové buněčné linie MeSH
• techniky dvojhybridového systému MeSH
• terciární struktura proteinů MeSH
• Check Tag
• lidé MeSH

Movement sequencing difficulties are part of the neurological soft signs (NSS), they have high clinical value because they are not always present in schizophrenia. We investigated the neuronal correlates of movement sequencing in 24 healthy controls and 24 schizophrenia patients, with (SZP SQ+) or without (SZP SQ-) sequencing difficulties. We characterized simultaneous and lagged functional connectivity between brain regions involved in movement sequencing using psychophysiological interaction (PPI) and the Granger causality modeling (GCM), respectively. Left premotor cortex (PMC) and superior parietal lobule (SPL) were specifically activated during sequential movements in all participants. Right PMC and precuneus, ipsilateral to the hand executing the task, activated during sequential movements only in healthy controls and SZP SQ-. SZP SQ+ showed hyperactivation in contralateral PMC, as compared to the other groups. PPI analysis revealed a deficit in inhibitory connections within this fronto-parietal network in SZP SQ+ during sequential task. GCM showed a significant lagged effective connectivity from right PMC to left SPL during task and rest periods in all groups and from right PMC to right precuneus in SZP SQ+ group only. Both SZP groups had a significant lagged connectivity from right to left PMC, during sequential task. Our results indicate that aberrant fronto-parietal network connectivity with cortical inhibition deficit and abnormal reliance on previous network activity are related to movement sequencing in SZP. The overactivation of motor cortex seems to be a good compensating strategy, the hyperactivation of parietal cortex is linked to motor deficit symptoms.

• MeSH
• dospělí MeSH
• funkční lateralita MeSH
• lidé MeSH
• lineární modely MeSH
• magnetická rezonanční tomografie MeSH
• mapování mozku * MeSH
• mladý dospělý MeSH
• mozek MeSH
• nervové dráhy MeSH
• počítačové zpracování obrazu MeSH
• pohybové poruchy etiologie   patologie   MeSH
• progrese nemoci MeSH
• psychomotorický výkon fyziologie   MeSH
• psychosomatické poruchy etiologie   MeSH
• schizofrenie komplikace   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

As with other drugs or pharmaceuticals, opioids differ in their rewarding or analgesic effects depending on when they are applied. In the previous study, we have demonstrated the day/night difference in the sensitivity of the major circadian clock in the suprachiasmatic nucleus to a low dose of morphine, and showed the bidirectional effect of morphine on pERK1/2 and pGSK3β levels in the suprachiasmatic nucleus depending on the time of administration. The main aim of this study was to identify other brain structures that respond differently to morphine depending on the time of its administration. Using immunohistochemistry, we identified 44 structures that show time-of-day specific changes in c-Fos level and activity of ERK1/2 and GSK3β kinases in response to a single dose of 1 mg/kg morphine. Furthermore, comparison among control groups revealed the differences in the spontaneous levels of all markers with a generally higher level during the night, that is, in the active phase of the day. We thus provide further evidence for diurnal variations in the activity of brain regions outside the suprachiasmatic nucleus indicated by the temporal changes in the molecular substrate. We suggest that these changes are responsible for generating diurnal variation in the reward behavior or analgesic effect of opioid administration.

• MeSH
• cirkadiánní rytmus fyziologie   MeSH
• kinasa glykogensynthasy 3beta metabolismus   MeSH
• krysa rodu rattus MeSH
• MAP kinasový signální systém fyziologie   MeSH
• morfin farmakologie   MeSH
• mozek účinky léků   metabolismus   MeSH
• opioidní analgetika farmakologie   MeSH
• potkani Wistar MeSH
• protoonkogenní proteiny c-fos metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The purpose of our study was to examine an early activation of JNK and p38 mitogen activated protein kinases (MAPK) and their substrate c-Myc after soman poisoning in order to enlighten the pathogenetic mechanism of nerve agent-induced non-specific effects. Male Wistar rats were intramuscularly poisoned by soman (60 μg.kg-1 - 70% LD50). Samples were taken 4, 24, and 72 hours after poisoning, immunohistochemically stained and phospho-JNKThr-183/Tyr-185, phospho-p38Thr180/Tyr182, and phospho-c- MycThr58/Ser62 expressions were measured using a computer Image analysis in apical and cryptal enterocytes of the colon transversum. We observed decreased phospho-JNK in apical enterocytes 4 and 24 h after poisoning and increased phospho-JNK in cryptal and apical enterocytes 72 h after intoxication. Phosphop38 dropped significantly in the apical compartment 72 h after soman poisoning. An activation of c-Myc decreased in both apical and cryptal compartment 4 and 24 h after soman intoxication, while increased in both compartments 72 h after poisoning. Soman poisoning seems to temporarily suppress promitotic pathways of proliferating cryptal cells and causes delayed activation of JNK stress signaling pathway.

• MeSH
• časové faktory MeSH
• colon transversum chemie   MeSH
• enterocyty chemie   MeSH
• experimenty na zvířatech MeSH
• financování organizované MeSH
• JNK mitogenem aktivované proteinkinasy chemie   MeSH
• kontrolní skupiny MeSH
• MAP kinasový signální systém fyziologie   MeSH
• mitogenem aktivované proteinkinasy p38 chemie   MeSH
• potkani Wistar MeSH
• protoonkogenní proteiny c-myc chemie   MeSH
• soman otrava   toxicita   MeSH
• výzkumný projekt MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH