title: The Effect of Surface Charge Saturation on Heat-induced Aggregation of Firefly Luciferase.
abstract: We present here the effect of firefly luciferase surface charge saturation and the presence of some additives on its thermal-induced aggregation. Three mutants of firefly luciferase prepared by introduction of surface Arg residues named as 2R, 3R and 5R have two, three and five additional arginine residues substituted at their surface compared to native luciferase; respectively. Turbidimetric study of heat-induced aggregation indicates that all three mutants were reproducibly aggregated at higher rates relative to wild type in spite of their higher thermostability. Among them, 2R had most evaluated propensity to heat-induced aggregation. Therefore, the hydrophilization followed by appearing of more substituted arginine residues with positive charge on the firefly luciferase surface was not reduced its thermal aggregation. Nevertheless, at the same condition in the presence of charged amino acids, e.g. Arg, Lys and Glu, as well as a hydrophobic amino acid, e.g. Val, the heat-induced aggregation of wild type and mutants of firefly luciferases was markedly decelerated than those in the absence of additives. On the basis of obtained results it seems, relinquishment of variety in charge of amino acid side chains, they via local interactions with proteins cause to decrease rate and extent of their thermal aggregation.
authors.affiliation: Department of Biology, Faculty of Basic Sciences, Tehran Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: Jamileh
authors.initials: J
authors.lastname: Gharanlar
authors.affiliation: Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Saman
authors.initials: S
authors.lastname: Hosseinkhani
authors.affiliation: Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Reza H
authors.initials: RH
authors.lastname: Sajedi
authors.affiliation: Department of Biology, Faculty of Basic Sciences, Tehran Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: Parichehr
authors.initials: P
authors.lastname: Yaghmaei
mesh_terms: D000818:Animals; D004795:Enzyme Stability; D048888:Fireflies / Q000201:enzymology*; D006358:Hot Temperature; D049409:Luciferases, Firefly / Q000737:chemistry / Q000378:metabolism*; D009391:Nephelometry and Turbidimetry; D013499:Surface Properties
chemical_list: D049409:Luciferases, Firefly