trifunctional
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Facultative anaerobic bacteria such as Escherichia coli have two α2β2 heterotetrameric trifunctional enzymes (TFE), catalyzing the last three steps of the β-oxidation cycle: soluble aerobic TFE (EcTFE) and membrane-associated anaerobic TFE (anEcTFE), closely related to the human mitochondrial TFE (HsTFE). The cryo-EM structure of anEcTFE and crystal structures of anEcTFE-α show that the overall assembly of anEcTFE and HsTFE is similar. However, their membrane-binding properties differ considerably. The shorter A5-H7 and H8 regions of anEcTFE-α result in weaker α-β as well as α-membrane interactions, respectively. The protruding H-H region of anEcTFE-β is therefore more critical for membrane-association. Mutational studies also show that this region is important for the stability of the anEcTFE-β dimer and anEcTFE heterotetramer. The fatty acyl tail binding tunnel of the anEcTFE-α hydratase domain, as in HsTFE-α, is wider than in EcTFE-α, accommodating longer fatty acyl tails, in good agreement with their respective substrate specificities.
- MeSH
- anaerobióza MeSH
- enoyl-CoA-hydratasa * chemie metabolismus MeSH
- Escherichia coli * genetika metabolismus MeSH
- lidé MeSH
- mitochondrie metabolismus MeSH
- oxidace-redukce MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Prezentujeme kazuistiku dvouletého chlapce s recidivujícími epizodami rhabdomyolýzy. Při metabolickém vyšetření v Ústavu dědičných metabolických poruch (ÚDMP) bylo u pacienta vysloveno podezření na poruchu beta-oxidace mastných kyselin (BOX MK) s dlouhým řetězcem. Výsledky dalších speciálních vyšetření na úrovni enzymů i genu vedly ke stanovení diagnózy deficitu mitochondriálního trifunkčního proteinu (MTP). V diskuzi jsou v přehledu uvedeny jednotlivé příčiny rhabdomyolýzy, její klinické projevy, diagnostický přístup i léčba.
A case report of 2-year-old boy with relapsing episodes of rhabdomyolysis is presented. The disorder of beta-oxidation of long chain fatty acid was presumed by the metabolic testing at the Institute of Inherited Metabolic Disorders. The results of further special testing on the level of enzymes and gene activities led to the diagnosis of the mitochondrial trifunctional protein (MTP) deficit. Various causes of rhabdomyolysis, its clinical manifestation, diagnostic approach and treatment are listed at a glance in the discussion.
- MeSH
- 3-hydroxyacyl-CoA dehydrogenasa mastných kyselin s dlouhým řetězcem nedostatek MeSH
- diferenciální diagnóza MeSH
- lidé MeSH
- mastné kyseliny metabolismus MeSH
- mitochondriální trifunkční protein nedostatek MeSH
- předškolní dítě MeSH
- rhabdomyolýza * diagnóza etiologie genetika metabolismus terapie MeSH
- vrozené poruchy metabolismu tuků * diagnóza komplikace patofyziologie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- předškolní dítě MeSH
- Publikační typ
- kazuistiky MeSH
- práce podpořená grantem MeSH
We designed a combinatorial library of trifunctional scaffold-derived compounds, which were derivatized with 30 different in-house-made azides. The compounds were proposed to mimic insulin receptor (IR)-binding epitopes in the insulin molecule and bind to and activate this receptor. This work has enabled us to test our synthetic and biological methodology and to prove its robustness and reliability for the solid-phase synthesis and testing of combinatorial libraries of the trifunctional scaffold-derived compounds. Our effort resulted in the discovery of two compounds, which were able to weakly induce the autophosphorylation of IR and weakly bind to this receptor at a 0.1 mM concentration. Despite these modest biological results, which well document the well-known difficulty in modulating protein-protein interactions, this study represents a unique example of targeting the IR with a set of nonpeptide compounds that were specifically designed and synthesized for this purpose. We believe that this work can open new perspectives for the development of next-generation insulin mimetics based on the scaffold structure.
- MeSH
- azidy chemická syntéza chemie MeSH
- inzulin analogy a deriváty chemie metabolismus MeSH
- knihovny malých molekul chemická syntéza chemie metabolismus farmakologie MeSH
- měď analýza MeSH
- molekulární struktura MeSH
- receptor inzulinu chemie metabolismus MeSH
- reprodukovatelnost výsledků MeSH
- techniky kombinatorické chemie * MeSH
- techniky syntézy na pevné fázi MeSH
- vazba proteinů MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
We present a trifunctional scaffold designed for the solid-phase synthesis of trimodal compounds. This scaffold holds two alkyne arms in a free and TIPS-protected form for consecutive CuAAC (copper(I)-catalyzed azide-alkyne cycloaddition), one Fmoc-protected hydrazide arm for reaction with aldehydes, and one carboxylic acid arm with CF₂ groups for attachment to the resin and (19)F-NMR quantification. This scaffold was attached to a resin and derivatized with model azides and aliphatic, electron-rich or electron-poor aromatic aldehydes. We identified several limitations of the scaffold caused by the instability of hydrazones in acidic conditions, in the presence of copper during CuAAC, and when copper accumulated in the resin. We successfully overcame these drawbacks by optimizing synthetic conditions for the derivatization of the scaffold with aromatic aldehydes. Overall, the new trifunctional scaffold combines CuAAC and hydrazone chemistries, offering a broader chemical space for the development of bioactive compounds.
Long chain 3-hydroxyacyl-CoA dehydrogenase deficiency (LCHADD/MTPD) and medium chain acyl-CoA dehydrogenase deficiency (MCADD) were included in the expanded neonatal screening program (ENBS) in Czechia in 2009, allowing for the presymptomatic diagnosis and nutritional management of these patients. The aim of our study was to assess the nationwide impact of ENBS on clinical outcome. This retrospective study analysed acute events and chronic complications and their severity in pre-ENBS and post-ENBS cohorts. In total, 28 children (12 before, 16 after ENBS) were diagnosed with LCHADD/MTPD (incidence 0.8/100,000 before and 1.2/100,000 after ENBS). In the subgroup detected by ENBS, a significantly longer interval from birth to first acute encephalopathy was observed. In addition, improvement in neuropathy and cardiomyopathy (although statistically non-significant) was demonstrated in the post-ENBS subgroup. In the MCADD cohort, we included 69 patients (15 before, 54 after ENBS). The estimated incidence rose from 0.7/100,000 before to 4.3/100,000 after ENBS. We confirmed a significant decrease in the number of episodes of acute encephalopathy and lower proportion of intellectual disability after ENBS (p < 0.0001). The genotype-phenotype correlations suggest a new association between homozygosity for the c.1528C > G variant and more severe heart involvement in LCHADD patients.
- MeSH
- 3-hydroxyacyl-CoA-dehydrogenasy nedostatek MeSH
- acyl-CoA-dehydrogenasa nedostatek MeSH
- dítě MeSH
- hodnocení výsledků zdravotní péče MeSH
- incidence MeSH
- kardiomyopatie diagnóza dietoterapie epidemiologie MeSH
- karnitin analogy a deriváty krev MeSH
- kojenec MeSH
- lidé MeSH
- mitochondriální myopatie diagnóza dietoterapie epidemiologie MeSH
- mitochondriální trifunkční protein nedostatek MeSH
- nemoci nervového systému diagnóza dietoterapie epidemiologie MeSH
- novorozenec MeSH
- novorozenecký screening metody MeSH
- předškolní dítě MeSH
- retrospektivní studie MeSH
- rhabdomyolýza diagnóza dietoterapie epidemiologie MeSH
- stupeň závažnosti nemoci MeSH
- vrozené poruchy metabolismu tuků diagnóza dietoterapie epidemiologie MeSH
- vrozené poruchy metabolismu diagnóza MeSH
- Check Tag
- dítě MeSH
- kojenec MeSH
- lidé MeSH
- mužské pohlaví MeSH
- novorozenec MeSH
- předškolní dítě MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
In humans, GART [phosphoribosylglycinamide formyltransferase (EC 2.1.2.2) / phosphoribosylglycinamide synthetase (EC 6.3.4.13) / phosphoribosylaminoimidazole synthetase (EC 6.3.3.1)] is a trifunctional protein which catalyzes the second, third, and fifth reactions of the ten step de novo purine synthesis (DNPS) pathway. The second step of DNPS is conversion of phosphoribosylamine (5-PRA) to glycineamide ribonucleotide (GAR). 5-PRA is extremely unstable under physiological conditions and is unlikely to accumulate in the absence of GART activity. Recently, a HeLa cell line null mutant for GART was constructed via CRISPR-Cas9 mutagenesis. This cell line, crGART, is an important cellular model of DNPS inactivation that does not accumulate DNPS pathway intermediates. In the current study, we characterized the crGART versus HeLa transcriptomes in purine-supplemented and purine-depleted growth conditions. We observed multiple transcriptome changes and discuss pathways and ontologies particularly relevant to Alzheimer disease and Down syndrome. We selected the Cluster of Differentiation (CD36) gene for initial analysis based on its elevated expression in crGART versus HeLa as well as its high basal expression, high log2 value, and minimal P-value.
Využití monoklonálních protilátek významně obohatilo možnosti léčby hematologických malignit i solidních nádorů. Rituximab v kombinaci s chemoterapií se stal standardním léčebným postupem u folikulárních lymfomů, chronické lymfocytární leukemie a difuzního velkobuněčného lymfomu. Znalost exprese povrchových antigenů na lymfomových buňkách umožnilo konstrukci většího počtu dalších monoklonálních protilátek. Je uvedena charakteristika používaných nebo zkoušených protilátek. Podobně jsou zaváděny monoklonální protilátky i k léčbě solidních nádorů. Jsou směrovány proti různým cílovým strukturám, jako jsou růstové faktory (např. bevacizumab) nebo proti extracelulární doméně epidermálního růstového faktoru (např. trastuzumab, cetuximab). Ke zvýšení terapeutické účinnosti byly připraveny konjugované protilátky. V klinickém použití nebo testování jsou protilátky konjugované s radionuklidem (90Y), s cytostatikem (calicheamycin) nebo s toxinem (estafenatox). Prohlubující se poznatky z oblasti molekulární a buněčné biologie umožnily vývoj nové generace protilátek (bispecifické/trifunkční protilátky, diabodies, peptibodies, intrabodies, imunocytokiny).
Monoclonal antibody-based therapeutic approaches have a significant impact in the treatment of hematologic malignancies and solid tumors. Rituximab in conjunction with chemotherapy is now standard treatment for follicular lymphoma, chronic lymphocytic leukemia and diffuse large B-cell lymphoma. Furthermore, based on our knowledge of antigen expression on the surface of lymphoma cells has led to development of the other monoclonal antibodies. Those, in clinical use or in clinical trials are shortly described. Similarly many monoclonal antibodies have been introduced for the treatment of solid tumors. These are directed against different targets such as growth factors (e.g. bevacizumab) or against the extracellular domain of epidermal growth factor (i.e. trastuzumab, cetuximab). For improving therapeutic efficacy, conjugated antibodies have been developed. Antibodies conjugated with radioisotopes (90Y), cytotoxic drugs (calicheamycin) or toxins (estafenatox) are used or are under investigation. Knowledge gained from molecular and cellular biology over past decades has driven the development of a new generation of monoclonal antibodies (bispecific/trifunctional antibodies, diabodies, peptibodies, intrabodies, immunocytokines).
- Klíčová slova
- mechanismus účinku, monospecifické protilátky, konjugované protilátky, protilátky nové generace,
- MeSH
- financování organizované MeSH
- lidé MeSH
- monoklonální protilátky farmakologie klasifikace terapeutické užití MeSH
- protilátky bispecifické farmakologie imunologie terapeutické užití MeSH
- protinádorové látky terapeutické užití MeSH
- specificita protilátek imunologie účinky léků MeSH
- Check Tag
- lidé MeSH
The new trinuclear tridentate Pt(II) complex [Pt(3)Cl(3)(hptab)](3+) (1; hptab = N,N,N',N',N'',N''-hexakis(2-pyridylmethyl)-1,3,5-tris(aminomethyl)benzene) exhibits promising cytotoxic effects in human and mouse tumor cells including those resistant to conventional cisplatin (Dalton Trans. 2006, 2617; Chem. Eur. J. 2009, 15, 5245). The present study is focused on the molecular pharmacology of 1, in particular on its interactions with DNA (which is the major pharmacological target of platinum antitumor drugs), to elucidate more deeply the mechanism underlying its antitumor effects. Results obtained with the aid of methods of molecular biophysics and pharmacology reveal new details of DNA modifications by 1. Complex 1 binds to DNA forming in the absence of proteins and molecular crowding agents mainly trifunctional intrastrand cross-links. In these DNA adducts all three Pt(II) centers of 1 are coordinated to DNA base residues, which leads to extensive conformational alterations in DNA. An intriguing aspect of the DNA-binding mode of this trinuclear Pt(II) complex 1 is that it can cross-link proteins to DNA. Even more interestingly, 1 can cross-link in the presence of molecular crowding agent, which mimics environmental conditions in cell nucleus, two DNA duplexes in a high yield--a feature observed for the first time for antitumor trinuclear platinum complexes. Thus, the concept for the design of agents capable of forming intramolecular tridentate DNA adducts, DNA-protein and interduplex DNA-DNA cross-links based on trinuclear tridentate Pt(II) complexes with semirigid aromatic linkers may result in new compounds which exhibit a variety of biological effects and can be also useful in nucleic acids research.
- MeSH
- DNA chemie MeSH
- lidé MeSH
- molekulární struktura MeSH
- myši MeSH
- organoplatinové sloučeniny chemie metabolismus MeSH
- protinádorové látky chemie metabolismus MeSH
- reagencia zkříženě vázaná chemie metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Metabolické funkcie peroxizómov zahŕňajú oxidáciu širokého spektra látok za prítomnosti kyslíka, pričom produkovaný peroxid vodíka môže byť degradovaný pomocou katalázy, respektíve využitý ďalej v peroxidázovej reakcii. Z hľadiska bunkovej patológie sú najvýznamnejšie procesy a a b oxidácie karboxylových kyselín, zvlášť významná je b-oxidácia karboxylových kyselín s veľmi dlhým reťazcom, ktorá prebieha výlučne v peroxizómoch. Mutácie peroxizómových génov spôsobujú závažné metabolické poruchy. V súčasnosti sú známe dve desiatky peroxizómových dedičných ochorení, ktoré sa rozdeľujú na generalizované (porucha biogenézy peroxizómov) a na izolované defekty jednotlivých peroxizómových enzýmov. Kombinovaná incidencia peroxizómových dedičných ochorení sa v západnej Európe odhaduje na 1:10 000. Všetky okrem X-viazanej adrenoleukodystrofie sa vyznačujú autozómovo-recesívnym typom dedičnosti. U generalizovaných ochorení, ako sú Zellwegerov syndróm, neonatálna adrenoleukodystrofia, infantilná Refsumova choroba, rhizomelická chondrodysplázia punctata vo fenotypovom prejave dominujú poškodenia nervového systému, pečene a obličiek. Väčšina pacientov zomiera do 1 roka, prežívanie viac ako 3 roky je výnimočné. X-adrenoleukodystrofia, pseudoneonatálna adrenoleukodystrofia, deficiencia trifun- kčného enzýmu, Refsumova choroba, primárna hyperoxalúria a akatalazémia sa vyznačujú deficitom jednotlivého enzýmu. Najčastejším peroxizómovým dedičným ochorením je X-adrenoleukodystrofia, ktorá ma viacero klinických fenotypov s prejavmi v detskom veku, ale aj klinicky menej závažnú formu, ktorá sa manifestuje až v dospelom veku – adrenomyeloneuropatiu. V diagnostike týchto ochorení sa využívajú biochemické, molekulárno-genetické, mor- fologické a imunochemické metódy, ktoré umožňujú nielen postnatálnu, ale aj prenatálnu diagnostiku.
Metabolic function of peroxisomes includes oxidation of wide spectrum of substances in the presence of oxygen. Hydrogen peroxide formed at the same time is either degraded by catalase or further utilized in peroxidative reactions. From the view of cellular pathology, the most important becomes alpha and beta-oxidation of carboxylic acids, particularly beta-oxidation of long-chain carboxylic acids, which undergoes selectively in peroxisomes. Mutations of peroxisomal genes result in serious metabolic disorders. At present about twenty hereditary peroxisomal diseases has been described. One group of them includes generalized forms (impairment of peroxisome biogenesis); diseases of other group result from isolated defects of individual peroxisomal enzymes. Combined incidence of peroxisomal hereditary disorders in the Western Europe is estimated to be 1:10 000. Beside the X-linked adrenoleukodystrophy, all others have the autosomal-recessive type of heredity. In phenotypic manifestation of generalized forms, as in the Zellweger syndrome, neonatal adrenoleukodystrophy, infantile Refsum disease, rhizomelic chondrodysplasia pun- ctata, an impairment of the central nervous system, liver, and kidney dominate. Most of the patients die within one year, survival period longer than three years becomes exceptional. X-adrenoleukodystrophy, pseudoneonatal adrenoleukodystrophy, trifunctional enzyme deficiency, Refsum disease, primary hyperoxaluria, acatalasemia result from the deficiency of a single enzyme. The most frequent peroxiosomal hereditary disease, the X-adrenoleukodys- trophy, has several clinical phenotypes, which most frequently manifest already in infants. The disease has also a clinically less serious form, which manifest only in adults – the adrenomyeloneuropathy. For the postnatal but also for the prenatal diagnostics, methods of biochemistry, molecular genetics, morphology, and immunocytochemistry are necessary.
- MeSH
- adrenoleukodystrofie genetika MeSH
- lidé MeSH
- peroxizomální poruchy diagnóza genetika MeSH
- peroxizomy metabolismus MeSH
- Refsumova nemoc genetika MeSH
- Zellwegerův syndrom genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
