GFP–Margatoxin, a Genetically Encoded Fluorescent Ligand to Probe Affinity of Kv1.3 Channel Blockers

International Journal of Molecular Sciences

Mikhail P. Kirpichnikov¹  Kristina R. Denisova¹  Dmitry A. Dolgikh¹  Nikita A. Orlov¹  Alexey V. Feofanov¹  Sergey A. Yakimov²  Elena A. Kryukova²  Oksana V. Nekrasova² 

[1] Faculty of Biology, Lomonosov Moscow State University, 119234 Moscow, Russia;Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997 Moscow, Russia;

关键词: Kv1.3 channel;    margatoxin;    GFP;    pore blocker;    fluorescent ligand;    affinity;   

DOI  :  10.3390/ijms23031724

来源: DOAJ

【 摘 要 】

Peptide pore blockers and their fluorescent derivatives are useful molecular probes to study the structure and functions of the voltage-gated potassium Kv1.3 channel, which is considered as a pharmacological target in the treatment of autoimmune and neurological disorders. We present Kv1.3 fluorescent ligand, GFP–MgTx, constructed on the basis of green fluorescent protein (GFP) and margatoxin (MgTx), the peptide, which is widely used in physiological studies of Kv1.3. Expression of the fluorescent ligand in E. coli cells resulted in correctly folded and functionally active GFP–MgTx with a yield of 30 mg per 1 L of culture. Complex of GFP–MgTx with the Kv1.3 binding site is reported to have the dissociation constant of 11 ± 2 nM. GFP–MgTx as a component of an analytical system based on the hybrid KcsA–Kv1.3 channel is shown to be applicable to recognize Kv1.3 pore blockers of peptide origin and to evaluate their affinities to Kv1.3. GFP–MgTx can be used in screening and pre-selection of Kv1.3 channel blockers as potential drug candidates.

【 授权许可】

Unknown