Search results for "denaturation"
showing 7 items of 107 documents
Stability and disassembly properties of human naïve Hsp60 and bacterial GroEL chaperonins.
2015
Human Hsp60 chaperonin and its bacterial homolog GroEL, in association with the corresponding co-chaperonins Hsp10 and GroES, constitute important chaperone systems promoting the proper folding of several mitochondrial proteins. Hsp60 is also currently described as a ubiquitous molecule with multiple roles both in health conditions and in several diseases. Naïve Hsp60 bearing the mitochondrial import signal has been recently demonstrated to present different oligomeric organizations with respect to GroEL, suggesting new possible physiological functions. Here we present a combined investigation with circular dichroism and small-angle X-ray scattering of structure, self-organization, and sta…
Synthesis of nucleoside and nucleotide conjugates of bile acids, and polymerase construction of bile acid-functionalized DNA.
2010
Aqueous Sonogashira cross-coupling reactions of 5-iodopyrimidine or 7-iodo-7-deazaadenine nucleosides with bile acid-derived terminal acetylenes linked via an ester or amide tether gave the corresponding bile acid–nucleoside conjugates. Analogous reactions of halogenated nucleoside triphosphates gave directly bile acid-modified dNTPs. Enzymatic incorporation of these modified nucleotides to DNA was successfully performed using Phusion polymerase for primer extension. One of the dNTPs (dCTP bearing cholic acid) was also efficient for PCR amplification.
Design, synthesis, DNA-binding and cytotoxicity evaluation of new potential combilexines
2002
Combilexines, compounds in which a DNA intercalator is linked to a minor groove binding component, interact with the DNA in a sequence specific manner to yield in most cases compounds with anticancer activity. A series of new compounds closely related to netropsin in which the two components were linked by an amide group was synthesised as potential combilexines. As some of these compounds showed cytotoxic activity in vitro, an attempt was made to rationalise their mechanism of action. The DNA binding characteristics of the carboxamides were evaluated by thermal denaturation experiments and by ethidium bromide displacement assay. Their ability to inhibit the topoisomerase I was also determi…
Cholesterol as stabilizer of the oxytocin receptor
2002
AbstractThe function of the oxytocin receptor system is strongly dependent on steroids as demonstrated by several physiological studies. One key element of this dependence on steroids may be the interaction of cholesterol and the oxytocin receptor. In this study, we show that cholesterol stabilizes the solubilized human oxytocin receptor against thermal inactivation and proteolytic degradation. In the absence of additional cholesterol, the soluble receptor inactivates within minutes. Maximal stabilization of the oxytocin receptor requires a continuous supply with cholesterol from a cholesterol-rich environment. A structure–activity analysis of various cholesterol analogues and their effect …
Heat-sensitive UV-Vis measurements for Phytochrome
2016
Fytokromit ovat punaista valoa aistivia proteiineja, jotka säätelevät suurta joukkoa solun toimintoja kasvi-, bakteeri- ja sienisoluissa. Nämä valoaktiiviset proteiinit muuntavat konformaatiotaan palautuvasti aktiivisen (Pfr) ja inaktiivisen (Pr) tilan välillä. Fytokromien aminohappojen väliset heikot vuorovaikutukset - etenkin vetysidokset - vaikuttavat lopulta laskostumiseen ja proteiinin lämpöstabiiliuteen. Fytokromit esiintyvät homodimeerisinä rakenteina, joiden monomeerit koostuvat useista domeeneista. Nämä jakautuvat N- terminaaliseen valosensitiiviseen moduuliin (PSM) ja C-terminaaliseen OutPut moduliin (OPM). PSM koostuu kolmesta domeenista (PAS-GAF ja PHY). Tässä työssä tutkittiin …
Titration of poly(dA-dT).poly(dA-dT) in solution at variable NaCl concentration
2004
CD and uv absorption data showed that high molecular weight poly(dA-dT) poly(dA-dT), at 298 K, undergoes an acid-induced transition from B-double helix to random coil in NaCl solutions of different concentrations, ranging from 0.005 to 0.600M. Similarly, titration of the polynucleotide with a strong base causes duplex-to-single strands transition. The base- and acid-induced transitions were both reversible by back-titration (with an acid or, respectively, with a base): the apparent pKa were the same in both directions. However, the number of protons per titratable site (adenine N1) required to reach half-denaturation was in great excess over the stoichiometric value; to a much larger extent…
MbCo in Saccharide Solid Amorphous Systems: A Combined FTIR and SAXS Study
2011
Saccharides, and in particular trehalose, are known for their efficiency in protecting biostructures against environmental stress [1], although the preservation mechanism is still debated. Experiments and simulations [2,3] on carboxy-myoglobin (MbCO) showed that the protein dynamics is highly inhibited in dry trehalose matrices, the inhibition being dependent on the water content. In these conditions, a mutual protein-matrix structural and dynamic influence is observed.Here we report a combined FTIR and SAXS study on MbCO embedded in dry amorphous matrices of trehalose and sucrose. FTIR measurements were performed at different protein/sugar ratios, focussing on the stretching band of the bo…