Photo Cross-Linking Probes Containing ϵ-N-Thioacyllysine and ϵ-N-Acyl-(δ-aza)lysine Residues

Research output: Contribution to journalJournal articlepeer-review

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Photo Cross-Linking Probes Containing ϵ-N-Thioacyllysine and ϵ-N-Acyl-(δ-aza)lysine Residues. / Bæk, Michael; Martín-Gago, Pablo; Laursen, Jonas S.; Madsen, Julie L.H.; Chakladar, Saswati; Olsen, Christian A.

In: Chemistry - A European Journal, Vol. 26, No. 17, 23.03.2020, p. 3862-3869.

Research output: Contribution to journalJournal articlepeer-review

Harvard

Bæk, M, Martín-Gago, P, Laursen, JS, Madsen, JLH, Chakladar, S & Olsen, CA 2020, 'Photo Cross-Linking Probes Containing ϵ-N-Thioacyllysine and ϵ-N-Acyl-(δ-aza)lysine Residues', Chemistry - A European Journal, vol. 26, no. 17, pp. 3862-3869. https://doi.org/10.1002/chem.201905338

APA

Bæk, M., Martín-Gago, P., Laursen, J. S., Madsen, J. L. H., Chakladar, S., & Olsen, C. A. (2020). Photo Cross-Linking Probes Containing ϵ-N-Thioacyllysine and ϵ-N-Acyl-(δ-aza)lysine Residues. Chemistry - A European Journal, 26(17), 3862-3869. https://doi.org/10.1002/chem.201905338

Vancouver

Bæk M, Martín-Gago P, Laursen JS, Madsen JLH, Chakladar S, Olsen CA. Photo Cross-Linking Probes Containing ϵ-N-Thioacyllysine and ϵ-N-Acyl-(δ-aza)lysine Residues. Chemistry - A European Journal. 2020 Mar 23;26(17):3862-3869. https://doi.org/10.1002/chem.201905338

Author

Bæk, Michael ; Martín-Gago, Pablo ; Laursen, Jonas S. ; Madsen, Julie L.H. ; Chakladar, Saswati ; Olsen, Christian A. / Photo Cross-Linking Probes Containing ϵ-N-Thioacyllysine and ϵ-N-Acyl-(δ-aza)lysine Residues. In: Chemistry - A European Journal. 2020 ; Vol. 26, No. 17. pp. 3862-3869.

Bibtex

@article{adcf487ea1fe451384e61ad0f50c2ab9,
title = "Photo Cross-Linking Probes Containing ϵ-N-Thioacyllysine and ϵ-N-Acyl-(δ-aza)lysine Residues",
abstract = "Posttranslational modifications (PTMs) are important in the regulation of protein function, trafficking, localization, and marking for degradation. This work describes the development of peptide activity/affinity-based probes for the discovery of proteins that recognize novel acyl-based PTMs on lysine residues in the proteome. The probes contain surrogates of ϵ-N-acyllysine by introduction of either hydrazide or thioamide functionalities to circumvent hydrolysis of the modification during the experiments. In addition to the modified PTMs, the developed chemotypes were analyzed with respect to the effect of peptide sequence. The photo cross-linking conditions and subsequent functionalization of the covalent adducts were systematically optimized by applying fluorophore labeling and gel electrophoresis (in-gel fluorescence measurements). Finally, selected probes, containing the ϵ-N-glutaryllysine and ϵ-N-myristoyllysine analogues, were successfully applied for the enrichment of native, endogenous proteins from cell lysate, recapitulating the expected interactions of SIRT5 and SIRT2, respectively. Interestingly, the latter mentioned was able to pull down two different splice variants of SIRT2, which has not been achieved with a covalent probe before. Based on this elaborate proof-of-concept study, we expect that the technology will have broad future applications for pairing of novel PTMs with the proteins that target them in the cell.",
keywords = "activity-based probes, affinity-based probes, histone deacetylases, photo-affinity labeling, posttranslational modifications, sirtuins",
author = "Michael B{\ae}k and Pablo Mart{\'i}n-Gago and Laursen, {Jonas S.} and Madsen, {Julie L.H.} and Saswati Chakladar and Olsen, {Christian A.}",
year = "2020",
month = mar,
day = "23",
doi = "10.1002/chem.201905338",
language = "English",
volume = "26",
pages = "3862--3869",
journal = "Chemistry: A European Journal",
issn = "0947-6539",
publisher = "Wiley - V C H Verlag GmbH & Co. KGaA",
number = "17",

}

RIS

TY - JOUR

T1 - Photo Cross-Linking Probes Containing ϵ-N-Thioacyllysine and ϵ-N-Acyl-(δ-aza)lysine Residues

AU - Bæk, Michael

AU - Martín-Gago, Pablo

AU - Laursen, Jonas S.

AU - Madsen, Julie L.H.

AU - Chakladar, Saswati

AU - Olsen, Christian A.

PY - 2020/3/23

Y1 - 2020/3/23

N2 - Posttranslational modifications (PTMs) are important in the regulation of protein function, trafficking, localization, and marking for degradation. This work describes the development of peptide activity/affinity-based probes for the discovery of proteins that recognize novel acyl-based PTMs on lysine residues in the proteome. The probes contain surrogates of ϵ-N-acyllysine by introduction of either hydrazide or thioamide functionalities to circumvent hydrolysis of the modification during the experiments. In addition to the modified PTMs, the developed chemotypes were analyzed with respect to the effect of peptide sequence. The photo cross-linking conditions and subsequent functionalization of the covalent adducts were systematically optimized by applying fluorophore labeling and gel electrophoresis (in-gel fluorescence measurements). Finally, selected probes, containing the ϵ-N-glutaryllysine and ϵ-N-myristoyllysine analogues, were successfully applied for the enrichment of native, endogenous proteins from cell lysate, recapitulating the expected interactions of SIRT5 and SIRT2, respectively. Interestingly, the latter mentioned was able to pull down two different splice variants of SIRT2, which has not been achieved with a covalent probe before. Based on this elaborate proof-of-concept study, we expect that the technology will have broad future applications for pairing of novel PTMs with the proteins that target them in the cell.

AB - Posttranslational modifications (PTMs) are important in the regulation of protein function, trafficking, localization, and marking for degradation. This work describes the development of peptide activity/affinity-based probes for the discovery of proteins that recognize novel acyl-based PTMs on lysine residues in the proteome. The probes contain surrogates of ϵ-N-acyllysine by introduction of either hydrazide or thioamide functionalities to circumvent hydrolysis of the modification during the experiments. In addition to the modified PTMs, the developed chemotypes were analyzed with respect to the effect of peptide sequence. The photo cross-linking conditions and subsequent functionalization of the covalent adducts were systematically optimized by applying fluorophore labeling and gel electrophoresis (in-gel fluorescence measurements). Finally, selected probes, containing the ϵ-N-glutaryllysine and ϵ-N-myristoyllysine analogues, were successfully applied for the enrichment of native, endogenous proteins from cell lysate, recapitulating the expected interactions of SIRT5 and SIRT2, respectively. Interestingly, the latter mentioned was able to pull down two different splice variants of SIRT2, which has not been achieved with a covalent probe before. Based on this elaborate proof-of-concept study, we expect that the technology will have broad future applications for pairing of novel PTMs with the proteins that target them in the cell.

KW - activity-based probes

KW - affinity-based probes

KW - histone deacetylases

KW - photo-affinity labeling

KW - posttranslational modifications

KW - sirtuins

UR - http://www.scopus.com/inward/record.url?scp=85081038396&partnerID=8YFLogxK

U2 - 10.1002/chem.201905338

DO - 10.1002/chem.201905338

M3 - Journal article

C2 - 31922630

AN - SCOPUS:85081038396

VL - 26

SP - 3862

EP - 3869

JO - Chemistry: A European Journal

JF - Chemistry: A European Journal

SN - 0947-6539

IS - 17

ER -

ID: 240981052