Protein hydrolysis
Dotaz
Zobrazit nápovědu
Mouse double minute 2 (MDM2) has a phosphorylation site within a lid motif at Ser17 whose phosphomimetic mutation to Asp17 stimulates MDM2-mediated polyubiquitination of p53. MDM2 lid deletion, but not Asp17 mutation, induced a blue shift in the λ(max) of intrinsic fluorescence derived from residues in the central domain including Trp235, Trp303, Trp323, and Trp329. This indicates that the Asp17 mutation does not alter the conformation of MDM2 surrounding the tryptophan residues. In addition, Phe235 mutation enhanced MDM2 binding to p53 but did not stimulate its ubiquitination function, thus uncoupling increases in p53 binding from its E3 ubiquitin ligase function. However, the Asp17 mutation in MDM2 stimulated its discharge of the UBCH5a-ubiquitin thioester adduct (UBCH5a is a ubiquitin-conjugating enzyme E2D 1 UBC4/5 homolog yeast). This stimulation of ubiquitin discharge from E2 was independent of the p53 substrate. There are now four known effects of the Asp17 mutation on MDM2: (i) it alters the conformation of the isolated N-terminus as defined by NMR; (ii) it induces increased thermostability of the isolated N-terminal domain; (iii) it stimulates the allosteric interaction of MDM2 with the DNA-binding domain of p53; and (iv) it stimulates a novel protein-protein interaction with the E2-ubiquitin complex in the absence of substrate p53 that, in turn, increases hydrolysis of the E2-ubiquitin thioester bond. These data also suggest a new strategy to disrupt MDM2 function by targeting the E2-ubiquitin discharge reaction.
- MeSH
- alosterická regulace MeSH
- aminokyselinové motivy MeSH
- bodová mutace * MeSH
- hydrolýza MeSH
- konformace proteinů MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádorový supresorový protein p53 chemie metabolismus MeSH
- polyubikvitin metabolismus MeSH
- protoonkogenní proteiny c-mdm2 chemie genetika metabolismus MeSH
- sekvence aminokyselin MeSH
- terciární struktura proteinů MeSH
- ubikvitin konjugující enzymy metabolismus MeSH
- ubikvitin metabolismus MeSH
- ubikvitinace MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Poultry feathers make up for as much as 8.5% of chicken weight and represent a considerable amount of almost pure keratin waste which is not being adequately utilized at the present time. The present study dealt with the processing of poultry feathers through a two-stage alkaline-enzymatic hydrolysis. In the first stage, feathers were mixed with a 0.1 or 0.3% KOH water solution in a 1 : 50 ratio and were incubated at 70°C for 24 h. After adjusting pH to 9, the effects examined in the second processing stage on the amount of degraded feathers were those of proteolytic enzyme additions (1-5%), time (4-8 h) and temperature (50-70°C). Processing feathers in 0.3% KOH and hydrolysing for 8 h in the second stage at 70°C with a 5% dose of enzyme (relative to dry feathers weight) produced approx. 91% degradation. Keratin hydrolysate is distinct for its high nitrogen content and reasonable inorganic solids level. Two-stage technology of alkaline-enzymatic hydrolysing of poultry feathers in an environment of 0.3% KOH achieves high efficiency under quite mild reaction conditions (temperature not exceeding 70°C with pH in a mildly alkaline region), and is feasible from an economic viewpoint. Keratin hydrolysate can find particular application in packaging technology (films, foils and encapsulates).
- MeSH
- drůbež MeSH
- hydrolýza MeSH
- keratiny chemie MeSH
- koncentrace vodíkových iontů MeSH
- odpadky - odstraňování metody MeSH
- peří chemie MeSH
- proteasy chemie MeSH
- průmyslový odpad analýza statistika a číselné údaje MeSH
- teplota MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
... Motifs of Protein Structure -- Few general principles emerged from the first protein structure -- The ... ... Membrane Proteins 201 -- Membrane proteins are difficult to crystallize 202 -- Bacteriorhodopsin contains ... ... mutations in the GTP-binding loops 226 -- The molecular basis of autophosphorylation of viral p21 226 -- Protein-protein ... ... Prediction, Engineering, and Design of -- Protein Structures 247 -- Prediction of protein structure from ... ... of protein molecules 269 -- Protein crystals are difficult to grow 270 -- X-ray sources are either monochromatic ...
xv, 302 stran : ilustrace ; 28 cm
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been recently introduced to many diagnostic microbiological laboratories. Besides the identification of bacteria and fungi, that technique provides a potentially useful tool for the detection of antimicrobial resistance, especially of that conferred by β-lactamases. Here, we describe an assay allowing a detection of meropenem hydrolysis in clinical isolates of Enterobacteriaceae, Pseudomonas spp., and Acinetobacter baumannii using MALDI-TOF MS. This method is able to confirm carbapenemases within 3 h. The results are important for proper and fast intervention to limit the spread of carbapenemase-producing bacteria and provide information for appropriate initial therapy of the infections caused by these microbes.
- MeSH
- Acinetobacter baumannii účinky léků enzymologie genetika izolace a purifikace MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální proteiny genetika metabolismus MeSH
- beta-laktamasy genetika metabolismus MeSH
- beta-laktamová rezistence genetika MeSH
- biotest * MeSH
- exprese genu MeSH
- hydrolýza MeSH
- infekce bakteriemi rodu Acinetobacter diagnóza farmakoterapie mikrobiologie MeSH
- lidé MeSH
- pseudomonádové infekce diagnóza farmakoterapie mikrobiologie MeSH
- Pseudomonas účinky léků enzymologie genetika izolace a purifikace MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- thienamyciny farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A comparison of a matrix-assisted laser desorption ionization-time of flight mass spectrometric (MALDI-TOF MS) meropenem hydrolysis assay with the Carba NP test showed that both methods exhibited low sensitivity (approximately 76%), mainly due to the false-negative results obtained with OXA-48-type producers. The addition of NH4HCO3 to the reaction buffer for the MALDI-TOF MS assay dramatically improved its sensitivity (98%). Automatic interpretation of the MALDI-TOF MS assay, using the MBT STAR-BL software, generally agreed with the results obtained after manual analysis. For the Carba NP test, spectrophotometric analysis found six additional carbapenemase producers.
- MeSH
- automatizované zpracování dat MeSH
- bakteriální proteiny analýza MeSH
- beta-laktamasy analýza MeSH
- hydrogenuhličitany * MeSH
- hydrolýza MeSH
- laboratorní automatizace MeSH
- lidé MeSH
- pufry MeSH
- senzitivita a specificita MeSH
- software MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- thienamyciny metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- MeSH
- acetylcholin metabolismus MeSH
- acetylcholinesterasa metabolismus MeSH
- acetylthiocholin metabolismus MeSH
- Electrophorus metabolismus MeSH
- hydrolýza MeSH
- katalýza MeSH
- kinetika MeSH
- koncentrace vodíkových iontů MeSH
- rybí proteiny metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
... Analysis of Intact Protein 9 -- V. Estimation of Free Amino Acids 20 -- VI. ... ... Methods of Protein Hydrolysis 32 -- VII. ... ... Are Peptides Intermediates in Protein Synthesis? 254 -- VII. ... ... Primary Structures of Specific Proteins 357 -- IV. ... ... Xi -- Chapter 6 -- Intramolecular Bonds in Proteins -- II. ...
2nd ed. 13, 665 s. : il., tab.
