Fusion protein for light control of BK channel and preparation method and application thereof

Abstract

The invention provides a fusion protein for light control of a BK channel. The fusion protein comprises a cell membrane anchoring sequence, a fluorescent protein, a photosensitive protein LOV2 structural domain and a BK potassium ion channel specific binding protein which are sequentially connected from an N terminal to a C terminal, wherein the cell membrane anchoring sequence, the fluorescent protein, the photosensitive protein LOV2 structural domain and the BK potassium ion channel specific binding protein are directly connected or connected through connecting peptides. The fusion protein provided by the invention can realize function regulation and control of an endogenous BK channel under the condition of blue light stimulation.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a fusion protein for light-regulated BK channels, a preparation method and application thereof. Background technique [0002] Ion channels are one of the foundations of life activities. There are a large number of ion channels on the cell membranes and various organelle membranes of eukaryotes and prokaryotes. Ion channels play an important role in maintaining the stability of the ion environment inside and outside the membrane, regulating the excitability of cells, and activating signaling pathways. One of the most important functions of ion channels is to regulate the electrical activity of cells, thereby affecting physiological activities such as neurotransmitter release, gland secretion, and muscle contraction. [0003] The large conductance calcium-activated potassium ion channel, referred to as BK channel (Big potassium), gets its name from its very large single-ch...

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Problems solved by technology

[0006] Existing genetically encoded optogenetic tools for regulating ion channel function, taking lumitoxin (literal translation: phototoxin) developed by Tillberg et al. in 2014 as an example (Schmidt, Tillberg, Chen and Boyden, A fully genetically encoded protein architecture for optical control of Peptideligand concentration, Nature Communications, 5, 3019, 2014), combine the light-sensitive protein LOV2 domain with the polypeptide toxin that specifically binds to the ion channel to block the ion channel and restore the opening of the ion channel under the action of blue light, However, when the tool is used, the ion channel is blocked in the absence of light, which has a certain impact on the normal physiological state of the cell.

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