Fluorescent Ligands on the Basis of Hongotoxin 1: eGFP-Hongotoxin 1

Savelieva, Maria; Kudryashova, Kseniya; Nekrasova, Oksana V.; Feofanov, Аlexey V.

doi:10.1017/s1431927619007049

Cited by 3 publications

(6 citation statements)

References 5 publications

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“…These K ap values for HgTx1 and ChTx correspond well with the values obtained for the same peptides using the KcsA-Kv1.3-based system with GFP-L1-HgTx1 as a fluorescent ligand: 200 and 115 pM, respectively [37]. Similar affinities were reported for HgTx1 and ChTx using other binding assays.…”

Section: Gfp-mgtx As a Fluorescent Probe To Recognize Kv13 Blockerssupporting

confidence: 86%

“…This seems to be a general property of GFP-tagged peptide blockers. Thus, dissociation constants of the complexes between KcsA-Kv1.3 and previously studied GFP-HgTx1, GFP-L2-AgTx2, His6-GFP-L2-AgTx2 and GFP-OSK1 were 1.1 ± 0.2, 16 ± 4, 10 ± 4, and 1.9 ± 0.4 nM, respectively [18,19,37]. Peptide blockers AgTx2, HgTx1, MgTx, and OSK1 derived from the scorpion venoms have picomolar affinities to Kv1.3 channel (https://kaliumdb.org/, accessed on 2 January 2022).…”

Section: Gfp-mgtx Interaction With the Kv13 Binding Sitementioning

confidence: 73%

“…The yield of GFP-MgTx was 30 mg per 1 L of cell culture that provides a significant amount of a fluorescent probe at a relatively low cost. At the same time, the achieved yield of the product is inferior to the yields of GFP-AgTx2 [19] and GFP-HgTx1 [37] (ca. 80 mg per 1 L) that may be due to more stringent conditions of Ni-chromatography for GFP-MgTx.…”

Section: Gfp-mgtx Interaction With the Kv13 Binding Sitementioning

confidence: 85%

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

Denisova

Orlov

Yakimov

et al. 2022

IJMS

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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.

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Section: Gfp-mgtx As a Fluorescent Probe To Recognize Kv13 Blockerssupporting

confidence: 86%

Section: Gfp-mgtx Interaction With the Kv13 Binding Sitementioning

confidence: 73%

Section: Gfp-mgtx Interaction With the Kv13 Binding Sitementioning

confidence: 85%

See 1 more Smart Citation

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

Denisova

Orlov

Yakimov

et al. 2022

IJMS

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“…1, F). It should be noted that Kd of RFP-HgTx1 is lower than that of previously studied eGFP-HgTx1 (3.2±1.4 nM for KcsA-Kv1.1 [5]), suggesting that a fluorescent protein tag is able to affect noticeably the blocker affinity to the target channel.…”

mentioning

confidence: 70%

“…An efficient alternative to chemical labeling was proposed by bioengineering of chimeric proteins that comprise peptide blockers fused to fluorescent proteins [4]. Recently, eGFP-HgTx1 chimera was obtained, that preserved high activity of HgTx1 for the target channels [5].…”

Section: Moskva Russiamentioning

confidence: 99%