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2. The effect of chemical modification by diethylpyrocarbonate on substrate and effector binding to phosphoenolpyruvate carboxylase from Crassula argentea. Evidence for a histidine residue in the malate binding site

Creator:
Meyer, C. R., Rustin, P., and Wedding, R. T.
Format:
print
Type:
model:internalpart and TEXT
Language:
Multiple languages
Description:
Chemical modification of purifíed phosphoenolpyravate carboxylase (PEPC) from the crassulacean acid metabolism plant Crassula argentea Thunb. with the histidyl reagent diethylpyrocarbonate (DEPC) resulted in a transient increase in activity followed by a decrease of activity with time. This biphasic response was observed when the modifíed enzyme was assayed at both low (sub-K,^ and saturating substráte (phosphoenolpyruvate, PEP) concentrations. There was an approximate 25- fold difference in the apparent rate constants for the activation and inhibition phases. This is in contrast to what we háve observed under similar conditions for the C4 enzyme from Zea mays L. for which only inhibition of activity occurs. Spectral studies indicated that up to 7 of the potential 20 histidine residues per subunit were modifíed, at least 3 of which were necessary for activity. The biphasic response of the Crassula enzyme was dependent on the concentration of DEPC. Progressively less inactivation was observed when modifying the enzyme with lower concentrations of DEPC. Chemical modification of PEPC with 75 pM DEPC resulted in a form of the enzyme with a lower K^, and higher This was concomitant with the modification of 4 histidines per subímit. Changes in the response of the enzyme to allosteric effectors were also observed; with modification the enzyme was desensitized to malate inhibition and glucose-6-phosphate activation. The Kj of the modifíed enzyme for malate increased over 15-fold. This was consistent with fluorescence binding studies using the extrinsic conformationalprobe S-anilino-l-naphthalenesulfonate which indicated the elimination of binding of malate and increased binding of the substráte to PEPC. Protection studies showed that malate desensitization was delayed by the presence of malate during modification. Malate also slowed the initial rate of histidine modifícation as measured spectrophotometrically. Thus histidine plays a role in the malate site of Crassula PEPC.
Rights:
http://creativecommons.org/licenses/by-nc-sa/4.0/ and policy:public

3. The influence of roughness on the discharge coefficient of a broad-crested weir

Creator:
Pařílková, Jana, Říha, Jaromír, and Zachoval, Zbyněk
Format:
bez média and svazek
Type:
model:article and TEXT
Subject:
levee, broad crested weir, discharge coefficient, roughness, ochranná hráz, přeliv se širokou korunou, součinitel průtoku, and drsnost
Language:
Slovak
Description:
The use of environmentally-friendly materials in hydraulic engineering (e.g. the stone lining of weirs at levees) calls for the more accurate estimation of the discharge coefficient for broad-crested weirs with a rough crest surface. However, in the available literature sources the discharge coefficient of broad-crested weirs is usually expressed for a smooth crest. The authors of this paper have summarized the theoretical knowledge related to the effect of weir crest surface roughness on the discharge coefficient. The method of determination of the head-discharge relation for broad-crested weirs with a rough crest surface is proposed based on known discharge coefficient values for smooth surfaces and on the roughness parameters of the weir. For selected scenarios the theoretical results were compared with experimental research carried out at the Laboratory of Water Management Research, Faculty of Civil Engineering (FCE), Brno University of Technology (BUT). and Používání přírodě blízkých materiálů ve vodním hospodářství (např. kamenná opevnění povrchu přelivů v ochranných hrázích) vyvolává požadavek přesnějšího stanovení součinitele průtoku pro přelivy se širokou a hydraulicky drsnou korunou. V dostupných literárních pramenech je součinitel průtoku přelivů se širokou korunou obvykle vyjádřen pouze pro hladký povrch koruny přelivu. Autoři článku shrnuli teoretické poznatky týkající se vlivu drsnosti povrchu koruny přelivu na součinitel průtoku a navrhli metodu pro stanovení způsobu hydraulického výpočtu konzumční křivky přepadu přes přeliv se širokou a drsnou korunou. Postup vychází ze znalosti součinitele průtoku přelivu s hladkou korunou a charakteristik drsnosti. Pro vybrané scénáře byly teoreticky vypočítané hodnoty porovnány s výsledky experimentálního výzkumu uskutečněného v Laboratoři vodohospodářského výzkumu Ústavu vodních staveb, Fakulty stavební, Vysokého učení technického v Brně.
Rights:
http://creativecommons.org/licenses/by-nc-sa/4.0/ and policy:public