Thermal stability of membrane-bound proteins
نویسندگان
چکیده
nanoDSF, the miniaturized differential scanning fluorimetry technology, is a revolutionary method to determine the thermostability of proteins by following changes in their intrinsic fluorescence. In this comparative study, the Prometheus NT.48 was used to determine the thermal stability of the membrane esterase PA2949 from Pseudomonas aeruginosa in presence of various detergents. The detergent type strongly affected enzyme thermal stability, which moreover correlated with enzyme activity. Thus, the Prometheus NT.48 can be used not only to rapidly screen for optimal purification conditions for enzymes, but also to evaluate enzyme activities based on their conformational stability in presence of detergents.
منابع مشابه
PRODUCTION, RELEASE AND THERMAL CHARACTERIZATION OF CELLULOLYTIC ENZYME FROM CELLULOMONAS sp. STRAIN "0"
Cellulase production by a Cellulomonas sp., isolated in 1985 from forest humus soil along the border of the Caspian Sea in Iran, was investigated. This strain secreted endo-and exo-cellulases in the culture medium, but some of the enzymes produced remained cell membrane bound. Cell bound enzymes were released by various treatments. The highest amount of endo-glucanase (up to 95%) and exo-gl...
متن کاملThe Extractability of Inner-Membrane Proteins from Salmonella typhimurium Intact Cells, Spheroplasts and Inner-Membrane Fragments by Non-Denaturing Detergents
The effect of Triton X-100, Na cholate and Tween 80 on the solubilization of integral membrane proteins in intact cells, spheroplasts and inner-membrane fragments of Salmonella typhimurium was studied. The detergents were used in various concentrations (1.6 to 64 mM) and cytochromes b and d were used as marker to monitor the solubilization of membrane-bound proteins. Results showed that no inne...
متن کاملProteases Detection of invitro Culture of Midgut Cells from Hyalomma anatolicum anatolicum (Acari: Ixodidae)
Proteases play a key role in protein digestion in ticks and other haematophagous insects. Our understanding of blood meal digestion in digestive system of ticks can be very useful for better understanding of basic rules for control of ticks. Cells of the midgut endocytose blood components. Blood proteins uptake by midgut cells, suggesting the presence of proteases in the midgut cells. In this...
متن کاملDisruption of a specific molecular interaction with a bound lipid affects the thermal stability of the purple bacterial reaction centre.
Relatively little is known about the functions of specific molecular interactions between membrane proteins and membrane lipids. The structural and functional consequences of disrupting a previously identified interaction between a molecule of the diacidic lipid cardiolipin and the purple bacterial reaction centre were examined. Mutagenesis of a highly conserved arginine (M267) that is responsi...
متن کاملThermal aggregation of egg white proteins as affected by saccharides
Thermal characteristics of egg white proteins (EWP) may differ in the presence of saccharides. Therefore, the influence of saccharides including carboxymethyl cellulose (CMC), pectin, sucrose and maltodextrin and heating time on physicochemical characteristics of EWP as a whole were studied. Investigation of Heat Coagulation Time (HCT), solubility, turbidity and protein secondary structure of h...
متن کامل