A soluble, folded protein without charged amino acid residues

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A soluble, folded protein without charged amino acid residues. / Højgaard, Casper; Kofoed, Christian; Espersen, Roall; Johansson, Kristoffer Enøe; Villa, Mara; Willemoës, Martin; Lindorff-Larsen, Kresten; Teilum, Kaare; Winther, Jakob R.

I: Biochemistry, Bind 55, Nr. 28, 2016, s. 3949–3956.

Publikation: Bidrag til tidsskriftTidsskriftartikelfagfællebedømt

Harvard

Højgaard, C, Kofoed, C, Espersen, R, Johansson, KE, Villa, M, Willemoës, M, Lindorff-Larsen, K, Teilum, K & Winther, JR 2016, 'A soluble, folded protein without charged amino acid residues', Biochemistry, bind 55, nr. 28, s. 3949–3956. https://doi.org/10.1021/acs.biochem.6b00269

APA

Højgaard, C., Kofoed, C., Espersen, R., Johansson, K. E., Villa, M., Willemoës, M., Lindorff-Larsen, K., Teilum, K., & Winther, J. R. (2016). A soluble, folded protein without charged amino acid residues. Biochemistry, 55(28), 3949–3956. https://doi.org/10.1021/acs.biochem.6b00269

Vancouver

Højgaard C, Kofoed C, Espersen R, Johansson KE, Villa M, Willemoës M o.a. A soluble, folded protein without charged amino acid residues. Biochemistry. 2016;55(28):3949–3956. https://doi.org/10.1021/acs.biochem.6b00269

Author

Højgaard, Casper ; Kofoed, Christian ; Espersen, Roall ; Johansson, Kristoffer Enøe ; Villa, Mara ; Willemoës, Martin ; Lindorff-Larsen, Kresten ; Teilum, Kaare ; Winther, Jakob R. / A soluble, folded protein without charged amino acid residues. I: Biochemistry. 2016 ; Bind 55, Nr. 28. s. 3949–3956.

Bibtex

@article{5465ce59813a485ab0fe4950057b97c6,
title = "A soluble, folded protein without charged amino acid residues",
abstract = "Charges are considered an integral part of protein structure and function, enhancing solubility and providing specificity in molecular interactions. We wished to investigate whether charged amino acids are indeed required for protein biogenesis and whether a protein completely free of titratable side chains can maintain solubility, stability, and function. As a model, we used a cellulose-binding domain from Cellulomonas fimi, which, among proteins of more than 100 amino acids, presently is the least charged in the Protein Data Bank, with a total of only four titratable residues. We find that the protein shows a surprising resilience toward extremes of pH, demonstrating stability and function (cellulose binding) in the pH range from 2 to 11. To ask whether the four charged residues present were required for these properties of this protein, we altered them to nontitratable ones. Remarkably, this chargeless protein is produced reasonably well in Escherichia coli, retains its stable three-dimensional structure, and is still capable of strong cellulose binding. To further deprive this protein of charges, we removed the N-terminal charge by acetylation and studied the protein at pH 2, where the C-terminus is effectively protonated. Under these conditions, the protein retains its function and proved to be both soluble and have a reversible folding-unfolding transition. To the best of our knowledge, this is the first time a soluble, functional protein with no titratable side chains has been produced.",
keywords = "Journal Article",
author = "Casper H{\o}jgaard and Christian Kofoed and Roall Espersen and Johansson, {Kristoffer En{\o}e} and Mara Villa and Martin Willemo{\"e}s and Kresten Lindorff-Larsen and Kaare Teilum and Winther, {Jakob R.}",
year = "2016",
doi = "10.1021/acs.biochem.6b00269",
language = "English",
volume = "55",
pages = "3949–3956",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "28",

}

RIS

TY - JOUR

T1 - A soluble, folded protein without charged amino acid residues

AU - Højgaard, Casper

AU - Kofoed, Christian

AU - Espersen, Roall

AU - Johansson, Kristoffer Enøe

AU - Villa, Mara

AU - Willemoës, Martin

AU - Lindorff-Larsen, Kresten

AU - Teilum, Kaare

AU - Winther, Jakob R.

PY - 2016

Y1 - 2016

N2 - Charges are considered an integral part of protein structure and function, enhancing solubility and providing specificity in molecular interactions. We wished to investigate whether charged amino acids are indeed required for protein biogenesis and whether a protein completely free of titratable side chains can maintain solubility, stability, and function. As a model, we used a cellulose-binding domain from Cellulomonas fimi, which, among proteins of more than 100 amino acids, presently is the least charged in the Protein Data Bank, with a total of only four titratable residues. We find that the protein shows a surprising resilience toward extremes of pH, demonstrating stability and function (cellulose binding) in the pH range from 2 to 11. To ask whether the four charged residues present were required for these properties of this protein, we altered them to nontitratable ones. Remarkably, this chargeless protein is produced reasonably well in Escherichia coli, retains its stable three-dimensional structure, and is still capable of strong cellulose binding. To further deprive this protein of charges, we removed the N-terminal charge by acetylation and studied the protein at pH 2, where the C-terminus is effectively protonated. Under these conditions, the protein retains its function and proved to be both soluble and have a reversible folding-unfolding transition. To the best of our knowledge, this is the first time a soluble, functional protein with no titratable side chains has been produced.

AB - Charges are considered an integral part of protein structure and function, enhancing solubility and providing specificity in molecular interactions. We wished to investigate whether charged amino acids are indeed required for protein biogenesis and whether a protein completely free of titratable side chains can maintain solubility, stability, and function. As a model, we used a cellulose-binding domain from Cellulomonas fimi, which, among proteins of more than 100 amino acids, presently is the least charged in the Protein Data Bank, with a total of only four titratable residues. We find that the protein shows a surprising resilience toward extremes of pH, demonstrating stability and function (cellulose binding) in the pH range from 2 to 11. To ask whether the four charged residues present were required for these properties of this protein, we altered them to nontitratable ones. Remarkably, this chargeless protein is produced reasonably well in Escherichia coli, retains its stable three-dimensional structure, and is still capable of strong cellulose binding. To further deprive this protein of charges, we removed the N-terminal charge by acetylation and studied the protein at pH 2, where the C-terminus is effectively protonated. Under these conditions, the protein retains its function and proved to be both soluble and have a reversible folding-unfolding transition. To the best of our knowledge, this is the first time a soluble, functional protein with no titratable side chains has been produced.

KW - Journal Article

U2 - 10.1021/acs.biochem.6b00269

DO - 10.1021/acs.biochem.6b00269

M3 - Journal article

C2 - 27307139

VL - 55

SP - 3949

EP - 3956

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 28

ER -

ID: 165879483