title: Thermal Stability of Dopamine Transporters.
abstract: The thermal stabilities of the rat and mouse dopamine transporter (DAT) proteins were studied within the temperature range of 0-37°C. The inactivation of the protein was followed by monitoring changes in radioligand-specific binding. We found that the process followed a rate equation with first-order kinetics and was characterized by having a single rate constant k inact. The activation energies (E a) that were calculated from the Arrhenius plots (ln k inact vs. 1/T) were 43 ± 5 and 45 ± 6 kJ/mol for the rat (rDAT) and mouse (mDAT) transporters, respectively, and 44 ± 7 kJ/mol for rDAT from PC-6.3 cell line. These E a values were similar to the E a values of thermal inactivation of the muscarinic receptor from rat brain cortex and to the thermal inactivation of other transmembrane proteins. However, all of these activation energy values were significantly lower than the E a values for soluble single-subunit proteins of similar size. These results therefore suggest that the thermal stability of transmembrane proteins may be governed to a significant extent by cell membrane properties and by interactions between the membrane components and the protein. In contrast, the stability of soluble proteins seems to be mostly governed by protein structure and size, which determine the sum of the stabilizing intramolecular interactions within the protein molecule. It is therefore not surprising that cell membrane properties and composition may have significant effects on the functional properties of transmembrane proteins.
authors: Chair of Organic Chemistry, Institute of Chemistry, University of Tartu, Ravila 14a, 50411, Tartu, Estonia, siim.kukk@ut.ee.
authors: Siim
authors: S
authors: Kukk
authors: Vladimir
authors: V
authors: Stepanov
authors: Jaak
authors: J
authors: Järv
mesh_terms: D000818:Animals; D002460:Cell Line; D050483:Dopamine Plasma Membrane Transport Proteins / Q000737:chemistry* / Q000378:metabolism; D006358:Hot Temperature; D051379:Mice; D055550:Protein Stability; D051381:Rats; D017208:Rats, Wistar
chemical_list: D050483:Dopamine Plasma Membrane Transport Proteins