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Promoting protein self-association in non-glycosylated Thermomyces lanuginosus lipase based on crystal lattice contacts

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Standard

Promoting protein self-association in non-glycosylated Thermomyces lanuginosus lipase based on crystal lattice contacts. / Madsen, Jens; Sørensen, Thomas Rebsdorf; Kaspersen, Jørn Døvling; Silow, Maria Berggård; Vind, Jesper; Pedersen, Jan Skov; Svendsen, Allan; Otzen, Daniel E.

In: BBA Proteins and Proteomics, Vol. 154, No. 12, 12.2015, p. 1914-1921.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Harvard

Madsen, J, Sørensen, TR, Kaspersen, JD, Silow, MB, Vind, J, Pedersen, JS, Svendsen, A & Otzen, DE 2015, 'Promoting protein self-association in non-glycosylated Thermomyces lanuginosus lipase based on crystal lattice contacts', BBA Proteins and Proteomics, vol. 154, no. 12, pp. 1914-1921. https://doi.org/10.1016/j.bbapap.2015.09.007

APA

Madsen, J., Sørensen, T. R., Kaspersen, J. D., Silow, M. B., Vind, J., Pedersen, J. S., Svendsen, A., & Otzen, D. E. (2015). Promoting protein self-association in non-glycosylated Thermomyces lanuginosus lipase based on crystal lattice contacts. BBA Proteins and Proteomics, 154(12), 1914-1921. https://doi.org/10.1016/j.bbapap.2015.09.007

CBE

Madsen J, Sørensen TR, Kaspersen JD, Silow MB, Vind J, Pedersen JS, Svendsen A, Otzen DE. 2015. Promoting protein self-association in non-glycosylated Thermomyces lanuginosus lipase based on crystal lattice contacts. BBA Proteins and Proteomics. 154(12):1914-1921. https://doi.org/10.1016/j.bbapap.2015.09.007

MLA

Vancouver

Madsen J, Sørensen TR, Kaspersen JD, Silow MB, Vind J, Pedersen JS et al. Promoting protein self-association in non-glycosylated Thermomyces lanuginosus lipase based on crystal lattice contacts. BBA Proteins and Proteomics. 2015 Dec;154(12):1914-1921. https://doi.org/10.1016/j.bbapap.2015.09.007

Author

Madsen, Jens ; Sørensen, Thomas Rebsdorf ; Kaspersen, Jørn Døvling ; Silow, Maria Berggård ; Vind, Jesper ; Pedersen, Jan Skov ; Svendsen, Allan ; Otzen, Daniel E. / Promoting protein self-association in non-glycosylated Thermomyces lanuginosus lipase based on crystal lattice contacts. In: BBA Proteins and Proteomics. 2015 ; Vol. 154, No. 12. pp. 1914-1921.

Bibtex

@article{1957e7cfa387424ca3f891298f774db0,
title = "Promoting protein self-association in non-glycosylated Thermomyces lanuginosus lipase based on crystal lattice contacts",
abstract = "We have used the crystal structure of Thermomyces lanuginosus lipase (TlL) to identify and strengthen potential protein-protein interaction sites in solution. As wildtype we used a deglycosylated mutant of TlL (N33Q). We designed a number of TlL mutants to promote interactions via interfaces detected in the crystal-lattice structure, through strengthening of hydrophobic, polar or electrostatic contacts or truncation of sterically blocking residues. We identify a mutant predicted to lead to increased interfacial hydrophobic contacts (N92F) that shows markedly increased self-association properties on native gradient gels. While wildtype TlL mainly forms monomer and <5% dimers, N92F forms stable trimers and dimers according to Size-Exclusion Chromatography and Small Angle X-ray Scattering. These oligomers account for ~25% of the population and their enzymatic activity is comparable to that of the monomer. Self-association stabilizes TlL against thermal denaturation. Furthermore, the trimer is stable to dilution and requires high concentrations (>2M) of urea to dissociate. We conclude that crystal lattice contacts are a good starting point for design strategies to promote protein self-association.",
author = "Jens Madsen and S{\o}rensen, {Thomas Rebsdorf} and Kaspersen, {J{\o}rn D{\o}vling} and Silow, {Maria Bergg{\aa}rd} and Jesper Vind and Pedersen, {Jan Skov} and Allan Svendsen and Otzen, {Daniel E}",
note = "Copyright {\textcopyright} 2015 Elsevier B.V. All rights reserved.",
year = "2015",
month = dec,
doi = "10.1016/j.bbapap.2015.09.007",
language = "English",
volume = "154",
pages = "1914--1921",
journal = "B B A - Proteins and Proteomics",
issn = "1570-9639",
publisher = "Elsevier BV",
number = "12",

}

RIS

TY - JOUR

T1 - Promoting protein self-association in non-glycosylated Thermomyces lanuginosus lipase based on crystal lattice contacts

AU - Madsen, Jens

AU - Sørensen, Thomas Rebsdorf

AU - Kaspersen, Jørn Døvling

AU - Silow, Maria Berggård

AU - Vind, Jesper

AU - Pedersen, Jan Skov

AU - Svendsen, Allan

AU - Otzen, Daniel E

N1 - Copyright © 2015 Elsevier B.V. All rights reserved.

PY - 2015/12

Y1 - 2015/12

N2 - We have used the crystal structure of Thermomyces lanuginosus lipase (TlL) to identify and strengthen potential protein-protein interaction sites in solution. As wildtype we used a deglycosylated mutant of TlL (N33Q). We designed a number of TlL mutants to promote interactions via interfaces detected in the crystal-lattice structure, through strengthening of hydrophobic, polar or electrostatic contacts or truncation of sterically blocking residues. We identify a mutant predicted to lead to increased interfacial hydrophobic contacts (N92F) that shows markedly increased self-association properties on native gradient gels. While wildtype TlL mainly forms monomer and <5% dimers, N92F forms stable trimers and dimers according to Size-Exclusion Chromatography and Small Angle X-ray Scattering. These oligomers account for ~25% of the population and their enzymatic activity is comparable to that of the monomer. Self-association stabilizes TlL against thermal denaturation. Furthermore, the trimer is stable to dilution and requires high concentrations (>2M) of urea to dissociate. We conclude that crystal lattice contacts are a good starting point for design strategies to promote protein self-association.

AB - We have used the crystal structure of Thermomyces lanuginosus lipase (TlL) to identify and strengthen potential protein-protein interaction sites in solution. As wildtype we used a deglycosylated mutant of TlL (N33Q). We designed a number of TlL mutants to promote interactions via interfaces detected in the crystal-lattice structure, through strengthening of hydrophobic, polar or electrostatic contacts or truncation of sterically blocking residues. We identify a mutant predicted to lead to increased interfacial hydrophobic contacts (N92F) that shows markedly increased self-association properties on native gradient gels. While wildtype TlL mainly forms monomer and <5% dimers, N92F forms stable trimers and dimers according to Size-Exclusion Chromatography and Small Angle X-ray Scattering. These oligomers account for ~25% of the population and their enzymatic activity is comparable to that of the monomer. Self-association stabilizes TlL against thermal denaturation. Furthermore, the trimer is stable to dilution and requires high concentrations (>2M) of urea to dissociate. We conclude that crystal lattice contacts are a good starting point for design strategies to promote protein self-association.

U2 - 10.1016/j.bbapap.2015.09.007

DO - 10.1016/j.bbapap.2015.09.007

M3 - Journal article

C2 - 26431886

VL - 154

SP - 1914

EP - 1921

JO - B B A - Proteins and Proteomics

JF - B B A - Proteins and Proteomics

SN - 1570-9639

IS - 12

ER -