Molecular Probes for Visualization of Nicotinic Acetylcholine Receptors Based on Snake Three-Finger Toxins and a Red Fluorescent Protein

Мұқаба

Дәйексөз келтіру

Толық мәтін

Ашық рұқсат Ашық рұқсат
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Рұқсат жабық Рұқсат ақылы немесе тек жазылушылар үшін

Аннотация

The visualization of macromolecular complexes is an important task in modern bioorganic chemistry, which can be addressed using various methodological approaches. The most widely used techniques involve radioactive- and fluorescent-labeled ligands. In this study, molecular probes were developed in the form of hybrid constructs combining one of three snake toxins (α-bungarotoxin, α-cobratoxin, or neurotoxin NT-II) with the red fluorescent protein mKate2. These chimeric proteins were produced in a bacterial expression system and purified via gel filtration. Using competitive radioligand binding assay with radiolabeled α-bungarotoxin, it was demonstrated that the obtained probes exhibit high affinity for the nicotinic acetylcholine receptor of the electric organ from orpedo californicaT, with half-maximal inhibitory concentration values in the nanomolar range. The fluorescent probes were successfully employed for the visualization of acetylcholine receptors on the surface of SH-SYSY cells.

Авторлар туралы

A. Kuzmenkov

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

Email: aleksey.kuzmenkov@gmail.com
Moscow, Russia

I. Chudetsky

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

Moscow, Russia

D. Kudryavtsev

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

Moscow, Russia

I. Kasheverov

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

Moscow, Russia

V. Tsetlin

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

Moscow, Russia

A. Vassilevski

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

Moscow, Russia

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