An ERG channel inhibitor from the scorpion Buthus eupeus

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Standard

An ERG channel inhibitor from the scorpion Buthus eupeus. / Korolkova, Y.V.; Kozlov, S.A.; Lipkin, A.V.; Pluzhnikov, K.A.; Hadley, J.K.; Filippov, A.K.; Brown, D.A.; Pedersen, Kamilla Angelo; Strøbaek, D.; Jespersen, Thomas; Olesen, Søren-Peter; Jensen Skanning, B.; Grishin, E.V.

I: Journal of Biological Chemistry, Bind 276, Nr. 13, 30.03.2001, s. 9868-76.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt

Harvard

Korolkova, YV, Kozlov, SA, Lipkin, AV, Pluzhnikov, KA, Hadley, JK, Filippov, AK, Brown, DA, Pedersen, KA, Strøbaek, D, Jespersen, T, Olesen, S-P, Jensen Skanning, B & Grishin, EV 2001, 'An ERG channel inhibitor from the scorpion Buthus eupeus', Journal of Biological Chemistry, bind 276, nr. 13, s. 9868-76. https://doi.org/10.1074/jbc.M005973200

APA

Korolkova, Y. V., Kozlov, S. A., Lipkin, A. V., Pluzhnikov, K. A., Hadley, J. K., Filippov, A. K., Brown, D. A., Pedersen, K. A., Strøbaek, D., Jespersen, T., Olesen, S-P., Jensen Skanning, B., & Grishin, E. V. (2001). An ERG channel inhibitor from the scorpion Buthus eupeus. Journal of Biological Chemistry, 276(13), 9868-76. https://doi.org/10.1074/jbc.M005973200

Vancouver

Korolkova YV, Kozlov SA, Lipkin AV, Pluzhnikov KA, Hadley JK, Filippov AK o.a. An ERG channel inhibitor from the scorpion Buthus eupeus. Journal of Biological Chemistry. 2001 mar. 30;276(13):9868-76. https://doi.org/10.1074/jbc.M005973200

Author

Korolkova, Y.V. ; Kozlov, S.A. ; Lipkin, A.V. ; Pluzhnikov, K.A. ; Hadley, J.K. ; Filippov, A.K. ; Brown, D.A. ; Pedersen, Kamilla Angelo ; Strøbaek, D. ; Jespersen, Thomas ; Olesen, Søren-Peter ; Jensen Skanning, B. ; Grishin, E.V. / An ERG channel inhibitor from the scorpion Buthus eupeus. I: Journal of Biological Chemistry. 2001 ; Bind 276, Nr. 13. s. 9868-76.

Bibtex

@article{45bdeb7074c711dbbee902004c4f4f50,
title = "An ERG channel inhibitor from the scorpion Buthus eupeus",
abstract = "The isolation of the peptide inhibitor of M-type K(+) current, BeKm-1, from the venom of the Central Asian scorpion Buthus eupeus has been described previously (Fillipov A. K., Kozlov, S. A., Pluzhnikov, K. A., Grishin, E. V., and Brown, D. A. (1996) FEBS Lett. 384, 277-280). Here we report the cloning, expression, and selectivity of BeKm-1. A full-length cDNA of 365 nucleotides encoding the precursor of BeKm-1 was isolated using the rapid amplification of cDNA ends polymerase chain reaction technique from mRNA obtained from scorpion telsons. Sequence analysis of the cDNA revealed that the precursor contains a signal peptide of 21 amino acid residues. The mature toxin consists of 36 amino acid residues. BeKm-1 belongs to the family of scorpion venom potassium channel blockers and represents a new subgroup of these toxins. The recombinant BeKm-1 was produced as a Protein A fusion product in the periplasm of Escherichia coli. After cleavage and high performance liquid chromatography purification, recombinant BeKm-1 displayed the same properties as the native toxin. Three BeKm-1 mutants (R27K, F32K, and R27K/F32K) were generated, purified, and characterized. Recombinant wild-type BeKm-1 and the three mutants partly inhibited the native M-like current in NG108-15 at 100 nm. The effect of the recombinant BeKm-1 on different K(+) channels was also studied. BeKm-1 inhibited hERG1 channels with an IC(50) of 3.3 nm, but had no effect at 100 nm on hEAG, hSK1, rSK2, hIK, hBK, KCNQ1/KCNE1, KCNQ2/KCNQ3, KCNQ4 channels, and minimal effect on rELK1. Thus, BeKm-1 was shown to be a novel specific blocker of hERG1 potassium channels.",
keywords = "Amino Acid Sequence, Animals, Base Sequence, Cation Transport Proteins, Cell Line, Chromatography, High Pressure Liquid, Cloning, Molecular, DNA, Complementary, DNA-Binding Proteins, Dose-Response Relationship, Drug, Electrophysiology, Escherichia coli, Ether-A-Go-Go Potassium Channels, Humans, Inhibitory Concentration 50, KCNQ Potassium Channels, KCNQ1 Potassium Channel, Kinetics, Mass Spectrometry, Mice, Molecular Sequence Data, Mutagenesis, Site-Directed, Mutation, Open Reading Frames, Polymerase Chain Reaction, Potassium Channel Blockers, Potassium Channels, Potassium Channels, Voltage-Gated, Protein Sorting Signals, Protein Structure, Tertiary, RNA, Messenger, Rats, Recombinant Fusion Proteins, Scorpion Venoms, Scorpions, Sequence Analysis, DNA, Sequence Homology, Amino Acid, Substrate Specificity, Time Factors, Trans-Activators, Tumor Cells, Cultured",
author = "Y.V. Korolkova and S.A. Kozlov and A.V. Lipkin and K.A. Pluzhnikov and J.K. Hadley and A.K. Filippov and D.A. Brown and Pedersen, {Kamilla Angelo} and D. Str{\o}baek and Thomas Jespersen and S{\o}ren-Peter Olesen and {Jensen Skanning}, B. and E.V. Grishin",
year = "2001",
month = mar,
day = "30",
doi = "10.1074/jbc.M005973200",
language = "English",
volume = "276",
pages = "9868--76",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology, Inc.",
number = "13",

}

RIS

TY - JOUR

T1 - An ERG channel inhibitor from the scorpion Buthus eupeus

AU - Korolkova, Y.V.

AU - Kozlov, S.A.

AU - Lipkin, A.V.

AU - Pluzhnikov, K.A.

AU - Hadley, J.K.

AU - Filippov, A.K.

AU - Brown, D.A.

AU - Pedersen, Kamilla Angelo

AU - Strøbaek, D.

AU - Jespersen, Thomas

AU - Olesen, Søren-Peter

AU - Jensen Skanning, B.

AU - Grishin, E.V.

PY - 2001/3/30

Y1 - 2001/3/30

N2 - The isolation of the peptide inhibitor of M-type K(+) current, BeKm-1, from the venom of the Central Asian scorpion Buthus eupeus has been described previously (Fillipov A. K., Kozlov, S. A., Pluzhnikov, K. A., Grishin, E. V., and Brown, D. A. (1996) FEBS Lett. 384, 277-280). Here we report the cloning, expression, and selectivity of BeKm-1. A full-length cDNA of 365 nucleotides encoding the precursor of BeKm-1 was isolated using the rapid amplification of cDNA ends polymerase chain reaction technique from mRNA obtained from scorpion telsons. Sequence analysis of the cDNA revealed that the precursor contains a signal peptide of 21 amino acid residues. The mature toxin consists of 36 amino acid residues. BeKm-1 belongs to the family of scorpion venom potassium channel blockers and represents a new subgroup of these toxins. The recombinant BeKm-1 was produced as a Protein A fusion product in the periplasm of Escherichia coli. After cleavage and high performance liquid chromatography purification, recombinant BeKm-1 displayed the same properties as the native toxin. Three BeKm-1 mutants (R27K, F32K, and R27K/F32K) were generated, purified, and characterized. Recombinant wild-type BeKm-1 and the three mutants partly inhibited the native M-like current in NG108-15 at 100 nm. The effect of the recombinant BeKm-1 on different K(+) channels was also studied. BeKm-1 inhibited hERG1 channels with an IC(50) of 3.3 nm, but had no effect at 100 nm on hEAG, hSK1, rSK2, hIK, hBK, KCNQ1/KCNE1, KCNQ2/KCNQ3, KCNQ4 channels, and minimal effect on rELK1. Thus, BeKm-1 was shown to be a novel specific blocker of hERG1 potassium channels.

AB - The isolation of the peptide inhibitor of M-type K(+) current, BeKm-1, from the venom of the Central Asian scorpion Buthus eupeus has been described previously (Fillipov A. K., Kozlov, S. A., Pluzhnikov, K. A., Grishin, E. V., and Brown, D. A. (1996) FEBS Lett. 384, 277-280). Here we report the cloning, expression, and selectivity of BeKm-1. A full-length cDNA of 365 nucleotides encoding the precursor of BeKm-1 was isolated using the rapid amplification of cDNA ends polymerase chain reaction technique from mRNA obtained from scorpion telsons. Sequence analysis of the cDNA revealed that the precursor contains a signal peptide of 21 amino acid residues. The mature toxin consists of 36 amino acid residues. BeKm-1 belongs to the family of scorpion venom potassium channel blockers and represents a new subgroup of these toxins. The recombinant BeKm-1 was produced as a Protein A fusion product in the periplasm of Escherichia coli. After cleavage and high performance liquid chromatography purification, recombinant BeKm-1 displayed the same properties as the native toxin. Three BeKm-1 mutants (R27K, F32K, and R27K/F32K) were generated, purified, and characterized. Recombinant wild-type BeKm-1 and the three mutants partly inhibited the native M-like current in NG108-15 at 100 nm. The effect of the recombinant BeKm-1 on different K(+) channels was also studied. BeKm-1 inhibited hERG1 channels with an IC(50) of 3.3 nm, but had no effect at 100 nm on hEAG, hSK1, rSK2, hIK, hBK, KCNQ1/KCNE1, KCNQ2/KCNQ3, KCNQ4 channels, and minimal effect on rELK1. Thus, BeKm-1 was shown to be a novel specific blocker of hERG1 potassium channels.

KW - Amino Acid Sequence

KW - Animals

KW - Base Sequence

KW - Cation Transport Proteins

KW - Cell Line

KW - Chromatography, High Pressure Liquid

KW - Cloning, Molecular

KW - DNA, Complementary

KW - DNA-Binding Proteins

KW - Dose-Response Relationship, Drug

KW - Electrophysiology

KW - Escherichia coli

KW - Ether-A-Go-Go Potassium Channels

KW - Humans

KW - Inhibitory Concentration 50

KW - KCNQ Potassium Channels

KW - KCNQ1 Potassium Channel

KW - Kinetics

KW - Mass Spectrometry

KW - Mice

KW - Molecular Sequence Data

KW - Mutagenesis, Site-Directed

KW - Mutation

KW - Open Reading Frames

KW - Polymerase Chain Reaction

KW - Potassium Channel Blockers

KW - Potassium Channels

KW - Potassium Channels, Voltage-Gated

KW - Protein Sorting Signals

KW - Protein Structure, Tertiary

KW - RNA, Messenger

KW - Rats

KW - Recombinant Fusion Proteins

KW - Scorpion Venoms

KW - Scorpions

KW - Sequence Analysis, DNA

KW - Sequence Homology, Amino Acid

KW - Substrate Specificity

KW - Time Factors

KW - Trans-Activators

KW - Tumor Cells, Cultured

U2 - 10.1074/jbc.M005973200

DO - 10.1074/jbc.M005973200

M3 - Journal article

C2 - 11136720

VL - 276

SP - 9868

EP - 9876

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 13

ER -

ID: 165653