Functional Significance of Conserved Cysteines in the Extracellular Loops of the ATP Binding Cassette Transporter Pdr11p

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Standard

Functional Significance of Conserved Cysteines in the Extracellular Loops of the ATP Binding Cassette Transporter Pdr11p. / Stanchev, Lyubomir Dimitrov; Marek, Magdalena; Xian, Feng; Klöhn, Mara; Silvestro, Daniele; Dittmar, Gunnar; López-Marqués, Rosa Laura; Günther Pomorski, Thomas.

In: Journal of Fungi, Vol. 7, No. 1, 2, 2021.

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

Harvard

Stanchev, LD, Marek, M, Xian, F, Klöhn, M, Silvestro, D, Dittmar, G, López-Marqués, RL & Günther Pomorski, T 2021, 'Functional Significance of Conserved Cysteines in the Extracellular Loops of the ATP Binding Cassette Transporter Pdr11p', Journal of Fungi, vol. 7, no. 1, 2. https://doi.org/10.3390/jof7010002

APA

Stanchev, L. D., Marek, M., Xian, F., Klöhn, M., Silvestro, D., Dittmar, G., López-Marqués, R. L., & Günther Pomorski, T. (2021). Functional Significance of Conserved Cysteines in the Extracellular Loops of the ATP Binding Cassette Transporter Pdr11p. Journal of Fungi, 7(1), [2]. https://doi.org/10.3390/jof7010002

Vancouver

Stanchev LD, Marek M, Xian F, Klöhn M, Silvestro D, Dittmar G et al. Functional Significance of Conserved Cysteines in the Extracellular Loops of the ATP Binding Cassette Transporter Pdr11p. Journal of Fungi. 2021;7(1). 2. https://doi.org/10.3390/jof7010002

Author

Stanchev, Lyubomir Dimitrov ; Marek, Magdalena ; Xian, Feng ; Klöhn, Mara ; Silvestro, Daniele ; Dittmar, Gunnar ; López-Marqués, Rosa Laura ; Günther Pomorski, Thomas. / Functional Significance of Conserved Cysteines in the Extracellular Loops of the ATP Binding Cassette Transporter Pdr11p. In: Journal of Fungi. 2021 ; Vol. 7, No. 1.

Bibtex

@article{398238592d96418f9f79043d5a95bf5f,
title = "Functional Significance of Conserved Cysteines in the Extracellular Loops of the ATP Binding Cassette Transporter Pdr11p",
abstract = "The pleiotropic drug resistance (PDR) transporter Pdr11p is expressed under anaerobic growth conditions at the plasma membrane of the yeast Saccharomyces cerevisiae, where it facilitates the uptake of exogenous sterols. Members of the fungal PDR family contain six conserved cysteines in their extracellular loops (ECL). For the functional analysis of these cysteine residues in Pdr11p, we generated a series of single cysteine-to-serine mutants. All mutant proteins expressed well and displayed robust ATPase activity upon purification. Mass-spectrometry analysis identified two cysteine residues (C582 and C603) in ECL3 forming a disulfide bond. Further characterization by cell-based assays showed that all mutants are compromised in facilitating sterol uptake, protein stability, and trafficking to the plasma membrane. Our data highlight the fundamental importance of all six extracellular cysteine residues for the functional integrity of Pdr11p and provide new structural insights into the PDR family of transporters.",
keywords = "ABC transport proteins, ATPase activity, Disulfide bonds, Protein trafficking, Sterol uptake",
author = "Stanchev, {Lyubomir Dimitrov} and Magdalena Marek and Feng Xian and Mara Kl{\"o}hn and Daniele Silvestro and Gunnar Dittmar and L{\'o}pez-Marqu{\'e}s, {Rosa Laura} and {G{\"u}nther Pomorski}, Thomas",
year = "2021",
doi = "10.3390/jof7010002",
language = "English",
volume = "7",
journal = "Journal of Fungi",
issn = "2309-608X",
publisher = "MDPI AG",
number = "1",

}

RIS

TY - JOUR

T1 - Functional Significance of Conserved Cysteines in the Extracellular Loops of the ATP Binding Cassette Transporter Pdr11p

AU - Stanchev, Lyubomir Dimitrov

AU - Marek, Magdalena

AU - Xian, Feng

AU - Klöhn, Mara

AU - Silvestro, Daniele

AU - Dittmar, Gunnar

AU - López-Marqués, Rosa Laura

AU - Günther Pomorski, Thomas

PY - 2021

Y1 - 2021

N2 - The pleiotropic drug resistance (PDR) transporter Pdr11p is expressed under anaerobic growth conditions at the plasma membrane of the yeast Saccharomyces cerevisiae, where it facilitates the uptake of exogenous sterols. Members of the fungal PDR family contain six conserved cysteines in their extracellular loops (ECL). For the functional analysis of these cysteine residues in Pdr11p, we generated a series of single cysteine-to-serine mutants. All mutant proteins expressed well and displayed robust ATPase activity upon purification. Mass-spectrometry analysis identified two cysteine residues (C582 and C603) in ECL3 forming a disulfide bond. Further characterization by cell-based assays showed that all mutants are compromised in facilitating sterol uptake, protein stability, and trafficking to the plasma membrane. Our data highlight the fundamental importance of all six extracellular cysteine residues for the functional integrity of Pdr11p and provide new structural insights into the PDR family of transporters.

AB - The pleiotropic drug resistance (PDR) transporter Pdr11p is expressed under anaerobic growth conditions at the plasma membrane of the yeast Saccharomyces cerevisiae, where it facilitates the uptake of exogenous sterols. Members of the fungal PDR family contain six conserved cysteines in their extracellular loops (ECL). For the functional analysis of these cysteine residues in Pdr11p, we generated a series of single cysteine-to-serine mutants. All mutant proteins expressed well and displayed robust ATPase activity upon purification. Mass-spectrometry analysis identified two cysteine residues (C582 and C603) in ECL3 forming a disulfide bond. Further characterization by cell-based assays showed that all mutants are compromised in facilitating sterol uptake, protein stability, and trafficking to the plasma membrane. Our data highlight the fundamental importance of all six extracellular cysteine residues for the functional integrity of Pdr11p and provide new structural insights into the PDR family of transporters.

KW - ABC transport proteins

KW - ATPase activity

KW - Disulfide bonds

KW - Protein trafficking

KW - Sterol uptake

U2 - 10.3390/jof7010002

DO - 10.3390/jof7010002

M3 - Journal article

C2 - 33375075

AN - SCOPUS:85098846375

VL - 7

JO - Journal of Fungi

JF - Journal of Fungi

SN - 2309-608X

IS - 1

M1 - 2

ER -

ID: 256935906