Title
Effects of his-tags on physical properties of parvalbumins
Author
Alisa A. Vologzhannikova, Polina A. Khorn, Alexei S. Kazakov, Eugene A. Permyakov, Vladimir N. Uversky, Sergei E. Permyakov
Year
2019
Journal
Cell Calcium
Abstract
A comparative study of His-tagged and non-tagged rat β-parvalbumin (rWT β-PA), calcium binding protein with the EF-hand calcium binding domains, has been carried out. The attachment of His-tag increases α-helical content and decreases β-sheets and β-turns content of the metal free form (apo-state) of β-PA. In contrast to this, the attachment of His-tag decreases α-helical content by more than 10% and increases contents of β-sheets and β-turns of the Ca2+-loaded state. According to the dynamic light scattering analysis, apo-state of His-tagged rat β-PA seems to be less compact compared with the apo-state of non-tagged rat β-PA. Surprisingly, the attachment of His-tag practically does not change mean hydrodynamic radius of Ca2+-loaded rat β-PA. The attachment of His-tag shifts thermal denaturation peaks of both apo- and Ca2+-loaded states of rat β-PA towards higher temperatures by 3–4 °C and slightly decreases its Ca2+ affinity. These results should be taken into consideration in the use of His-tagged parvalbumins.
Instrument
J-810
Keywords
Circular dichroism, Secondary structure, Chemical stability, Biochemistry