Properties of Potassium Channel Kv1.1 on the Basis of Fluorescent Dimer of Alpha-Subunits mKate2-Kv1.1-Kv1.1 in Neuro-2a Cells

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Abstract

The properties of the voltage-gated potassium Kv1.1 channel, formed in Neuro-2a cells from dimers of the human α-subunits Kv1.1 linked by the Lys-Leu dipeptide and fused at the N-terminus with the mKate2 fluorescent protein (mKate2-(Kv1.1)2), were studied. It was found that the linking of α-subunits Kv1.1 into a dimer did not affect the membrane expression of the channel and the features of its cellular distribution compared with monomers mKate2-Kv1.1. No differences were found between channels based on mKate2-(Kv1.1)2 dimers and mKate2-Kv1.1 monomers in the half-activation potential, channel activation constants, the magnitude and nature of potassium ion currents. The data obtained suggest the possibility of creating bioengineered protein structures by linking two different α-subunits in a similar way, which would form fluorescent heterotetrameric voltage-gated potassium channels with an α-subunit stoichiometry of 2 : 2 in mammalian cells.

About the authors

A. V Efremenko

Shenyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences

Moscow, Russia

E. V Kryukova

Shenyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences

Moscow, Russia

O. V Kazakov

Shenyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences

Moscow, Russia

M. P Kirpichnikov

Shenyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences; Lomonosov Moscow State University, Faculty of Biology

Moscow, Russia; Moscow, Russia

O. V Nekrasova

Shenyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences

Moscow, Russia

A. V Feofanov

Shenyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences; Lomonosov Moscow State University, Faculty of Biology

Email: avfeofanov@yandex.ru
Moscow, Russia; Moscow, Russia

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