Purification and initial characterization of Plasmodium falciparum K+ channels, PfKch1 and PfKch2 produced in Saccharomyces cerevisiae

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Purification and initial characterization of Plasmodium falciparum K+ channels, PfKch1 and PfKch2 produced in Saccharomyces cerevisiae. / Molbaek, Karen; Tejada, Maria; Ricke, Christina Hoeier; Scharff-Poulsen, Peter; Ellekvist, Peter; Helix-Nielsen, Claus; Kumar, Nirbhay; Klaerke, Dan A.; Pedersen, Per Amstrup.

In: Microbial Cell Factories, Vol. 19, No. 1, 183, 2020.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Molbaek, K, Tejada, M, Ricke, CH, Scharff-Poulsen, P, Ellekvist, P, Helix-Nielsen, C, Kumar, N, Klaerke, DA & Pedersen, PA 2020, 'Purification and initial characterization of Plasmodium falciparum K+ channels, PfKch1 and PfKch2 produced in Saccharomyces cerevisiae', Microbial Cell Factories, vol. 19, no. 1, 183. https://doi.org/10.1186/s12934-020-01437-7

APA

Molbaek, K., Tejada, M., Ricke, C. H., Scharff-Poulsen, P., Ellekvist, P., Helix-Nielsen, C., Kumar, N., Klaerke, D. A., & Pedersen, P. A. (2020). Purification and initial characterization of Plasmodium falciparum K+ channels, PfKch1 and PfKch2 produced in Saccharomyces cerevisiae. Microbial Cell Factories, 19(1), [183]. https://doi.org/10.1186/s12934-020-01437-7

Vancouver

Molbaek K, Tejada M, Ricke CH, Scharff-Poulsen P, Ellekvist P, Helix-Nielsen C et al. Purification and initial characterization of Plasmodium falciparum K+ channels, PfKch1 and PfKch2 produced in Saccharomyces cerevisiae. Microbial Cell Factories. 2020;19(1). 183. https://doi.org/10.1186/s12934-020-01437-7

Author

Molbaek, Karen ; Tejada, Maria ; Ricke, Christina Hoeier ; Scharff-Poulsen, Peter ; Ellekvist, Peter ; Helix-Nielsen, Claus ; Kumar, Nirbhay ; Klaerke, Dan A. ; Pedersen, Per Amstrup. / Purification and initial characterization of Plasmodium falciparum K+ channels, PfKch1 and PfKch2 produced in Saccharomyces cerevisiae. In: Microbial Cell Factories. 2020 ; Vol. 19, No. 1.

Bibtex

@article{43c4c19aec384a63b27ef94ef49170ee,
title = "Purification and initial characterization of Plasmodium falciparum K+ channels, PfKch1 and PfKch2 produced in Saccharomyces cerevisiae",
abstract = "Resistance towards known antimalarial drugs poses a significant problem, urging for novel drugs that target vital proteins in the malaria parasite Plasmodium falciparum. However, recombinant production of malaria proteins is notoriously difficult. To address this, we have investigated two putative K+ channels, PfKch1 and PfKch2, identified in the P. falciparum genome. We show that PfKch1 and PfKch2 and a C-terminally truncated version of PfKch1 (PfKch11-1094) could indeed be functionally expressed in vivo, since a K+-uptake deficient Saccharomyces cerevisiae strain was complemented by the P. falciparum cDNAs. PfKch11-1094-GFP and GFP-PfKch2 fusion proteins were overexpressed in yeast, purified and reconstituted in lipid bilayers to determine their electrophysiological activity. Single channel conductance amounted to 16 ± 1 pS for PfKch11-1094-GFP and 28 ± 2 pS for GFP-PfKch2. We predicted regulator of K+-conductance (RCK) domains in the C-terminals of both channels, and we accordingly measured channel activity in the presence of Ca2+.",
keywords = "K-channels, Malaria, Recombinant protein, Yeast",
author = "Karen Molbaek and Maria Tejada and Ricke, {Christina Hoeier} and Peter Scharff-Poulsen and Peter Ellekvist and Claus Helix-Nielsen and Nirbhay Kumar and Klaerke, {Dan A.} and Pedersen, {Per Amstrup}",
year = "2020",
doi = "10.1186/s12934-020-01437-7",
language = "English",
volume = "19",
journal = "Microbial Cell",
issn = "1475-2859",
publisher = "BioMed Central",
number = "1",

}

RIS

TY - JOUR

T1 - Purification and initial characterization of Plasmodium falciparum K+ channels, PfKch1 and PfKch2 produced in Saccharomyces cerevisiae

AU - Molbaek, Karen

AU - Tejada, Maria

AU - Ricke, Christina Hoeier

AU - Scharff-Poulsen, Peter

AU - Ellekvist, Peter

AU - Helix-Nielsen, Claus

AU - Kumar, Nirbhay

AU - Klaerke, Dan A.

AU - Pedersen, Per Amstrup

PY - 2020

Y1 - 2020

N2 - Resistance towards known antimalarial drugs poses a significant problem, urging for novel drugs that target vital proteins in the malaria parasite Plasmodium falciparum. However, recombinant production of malaria proteins is notoriously difficult. To address this, we have investigated two putative K+ channels, PfKch1 and PfKch2, identified in the P. falciparum genome. We show that PfKch1 and PfKch2 and a C-terminally truncated version of PfKch1 (PfKch11-1094) could indeed be functionally expressed in vivo, since a K+-uptake deficient Saccharomyces cerevisiae strain was complemented by the P. falciparum cDNAs. PfKch11-1094-GFP and GFP-PfKch2 fusion proteins were overexpressed in yeast, purified and reconstituted in lipid bilayers to determine their electrophysiological activity. Single channel conductance amounted to 16 ± 1 pS for PfKch11-1094-GFP and 28 ± 2 pS for GFP-PfKch2. We predicted regulator of K+-conductance (RCK) domains in the C-terminals of both channels, and we accordingly measured channel activity in the presence of Ca2+.

AB - Resistance towards known antimalarial drugs poses a significant problem, urging for novel drugs that target vital proteins in the malaria parasite Plasmodium falciparum. However, recombinant production of malaria proteins is notoriously difficult. To address this, we have investigated two putative K+ channels, PfKch1 and PfKch2, identified in the P. falciparum genome. We show that PfKch1 and PfKch2 and a C-terminally truncated version of PfKch1 (PfKch11-1094) could indeed be functionally expressed in vivo, since a K+-uptake deficient Saccharomyces cerevisiae strain was complemented by the P. falciparum cDNAs. PfKch11-1094-GFP and GFP-PfKch2 fusion proteins were overexpressed in yeast, purified and reconstituted in lipid bilayers to determine their electrophysiological activity. Single channel conductance amounted to 16 ± 1 pS for PfKch11-1094-GFP and 28 ± 2 pS for GFP-PfKch2. We predicted regulator of K+-conductance (RCK) domains in the C-terminals of both channels, and we accordingly measured channel activity in the presence of Ca2+.

KW - K-channels

KW - Malaria

KW - Recombinant protein

KW - Yeast

U2 - 10.1186/s12934-020-01437-7

DO - 10.1186/s12934-020-01437-7

M3 - Journal article

C2 - 32957994

AN - SCOPUS:85091475087

VL - 19

JO - Microbial Cell

JF - Microbial Cell

SN - 1475-2859

IS - 1

M1 - 183

ER -

ID: 249426160