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Hedberg, Christian
Alternative names
Publications (10 of 18) Show all publications
Ochtrop, P., Ernst, S., Itzen, A. & Hedberg, C. (2019). Exploring the Substrate Scope of the Bacterial Phosphocholine Transferase AnkX for Versatile Protein Functionalization. ChemBioChem (Print), 20(18), 2336-2340
Open this publication in new window or tab >>Exploring the Substrate Scope of the Bacterial Phosphocholine Transferase AnkX for Versatile Protein Functionalization
2019 (English)In: ChemBioChem (Print), ISSN 1439-4227, E-ISSN 1439-7633, Vol. 20, no 18, p. 2336-2340Article in journal (Refereed) Published
Abstract [en]

Site-specific protein functionalization has become an indispensable tool in modern life sciences. Here, tag-based enzymatic protein functionalization techniques are among the most versatilely applicable approaches. However, many chemo-enzymatic functionalization strategies suffer from low substrate scopes of the enzymes utilized for functional labeling probes. We report on the wide substrate scope of the bacterial enzyme AnkX towards derivatized CDP-choline analogues and demonstrate that AnkX-catalyzed phosphocholination can be used for site-specific one- and two-step protein labeling with a broad array of different functionalities, displaying fast second-order transfer rates of 5x10(2) to 1.8x10(4) m(-1) s(-1). Furthermore, we also present a strategy for the site-specific dual labeling of proteins of interest, based on the exploitation of AnkX and the delabeling function of the enzyme Lem3. Our results contribute to the wide field of protein functionalization, offering an attractive chemo-enzymatic tag-based modification strategy for in vitro labeling.

Place, publisher, year, edition, pages
Wiley-VCH Verlagsgesellschaft, 2019
Keywords
CDP-choline, dual labeling, phosphocholination, protein modifications, transferases
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:umu:diva-164139 (URN)10.1002/cbic.201900200 (DOI)000486565800006 ()31054261 (PubMedID)
Available from: 2019-10-17 Created: 2019-10-17 Last updated: 2019-10-17Bibliographically approved
diva2:1295208
Open this publication in new window or tab >>Photoactivated Colibactin Probes Induce Cellular DNA Damage
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2019 (English)In: Angewandte Chemie International Edition, ISSN 1433-7851, E-ISSN 1521-3773, Vol. 58, no 5, p. 1417-1421Article in journal (Refereed) Published
Abstract [en]

Colibactin is a small molecule produced by certain bacterial species of the human microbiota that harbour the pks genomic island. Pks(+) bacteria induce a genotoxic phenotype in eukaryotic cells and have been linked with colorectal cancer progression. Colibactin is produced in a benign, prodrug form which, prior to export, is enzymatically matured by the producing bacteria to its active form. Although the complete structure of colibactin has not been determined, key structural features have been described including an electrophilic cyclopropane motif, which is believed to alkylate DNA. To investigate the influence of the putative "warhead" and the prodrug strategy on genotoxicity, a series of photolabile colibactin probes were prepared that upon irradiation induced a pks(+) like phenotype in HeLa cells. Furthermore, results from DNA cross-linking and imaging studies of clickable analogues enforce the hypothesis that colibactin effects its genotoxicity by directly targeting DNA.

Keywords
click chemistry, colibactin, DNA damage, microbiome, photochemistry
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:umu:diva-156892 (URN)10.1002/anie.201812326 (DOI)000458826100026 ()30506956 (PubMedID)
Available from: 2019-03-11 Created: 2019-03-11 Last updated: 2019-03-11Bibliographically approved
Wolf-Watz, M., Rogne, P., Sauer-Eriksson, A. E., Sauer, U. H. & Hedberg, C. (2019). Positive and Negative Substrate Interference Supported by Coinciding Enzyme Residues. Paper presented at 63rd Annual Meeting of the Biophysical-Society, MAR 02-06, 2019, Baltimore, MD. Biophysical Journal, 116(3), 485A-485A
Open this publication in new window or tab >>Positive and Negative Substrate Interference Supported by Coinciding Enzyme Residues
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2019 (English)In: Biophysical Journal, ISSN 0006-3495, E-ISSN 1542-0086, Vol. 116, no 3, p. 485A-485AArticle in journal, Meeting abstract (Other academic) Published
Place, publisher, year, edition, pages
CELL PRESS, 2019
National Category
Biophysics
Identifiers
urn:nbn:se:umu:diva-157776 (URN)10.1016/j.bpj.2018.11.2620 (DOI)000460779802439 ()
Conference
63rd Annual Meeting of the Biophysical-Society, MAR 02-06, 2019, Baltimore, MD
Available from: 2019-04-10 Created: 2019-04-10 Last updated: 2019-04-10Bibliographically approved
Paulsen, M. H., Karlsen, E. A., Ausbacher, D., Anderssen, T., Bayer, A., Ochtrop, P., . . . Strøm, M. B. (2018). An amphipathic cyclic tetrapeptide scaffold containing halogenated β2,2-amino acids with activity against multiresistant bacteria. Journal of Peptide Science, 24(10), Article ID UNSP e3117.
Open this publication in new window or tab >>An amphipathic cyclic tetrapeptide scaffold containing halogenated β2,2-amino acids with activity against multiresistant bacteria
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2018 (English)In: Journal of Peptide Science, ISSN 1075-2617, E-ISSN 1099-1387, Vol. 24, no 10, article id UNSP e3117Article in journal (Refereed) Published
Abstract [en]

The present study describes the synthesis and biological studies of a small series of head-to-tail cyclic tetrapeptides of the general structure c(Lys‐β2,2‐Xaa‐Lys) containing one lipophilic β2,2-amino acid and Lys, Gly, Ala, or Phe as the Xaa residue in the sequence. The peptides were investigated for antimicrobial activity against gram-positive and gram-negative reference strains and 30 multiresistant clinical isolates including strains with extended spectrum β-lactamase-carbapenemase (ESBL-CARBA) production. Toxicity was determined against human red blood cells. The most potent peptides showed high activity against the gram-positive clinical isolates with minimum inhibitory concentrations of 4-8μg/mL and low haemolytic activity. The combination of high antimicrobial activity and low toxicity shows that these cyclic tetrapeptides containing lipophilic β2,2-amino acids form a valuable scaffold for designing novel antimicrobial agents.

Place, publisher, year, edition, pages
John Wiley & Sons, 2018
Keywords
antimicrobial peptides, beta-amino acids, CARBA, cyclic tetrapeptides, ESBL, MRSA, multiresistant bacteria, peptidomimetics, SMAMPs, synthetic mimics of antimicrobial peptides
National Category
Medicinal Chemistry
Identifiers
urn:nbn:se:umu:diva-153717 (URN)10.1002/psc.3117 (DOI)000445732400005 ()30112781 (PubMedID)
Available from: 2018-11-27 Created: 2018-11-27 Last updated: 2018-11-27Bibliographically approved
Rogne, P., Rosselin, M., Grundström, C., Hedberg, C., H. Sauer, U. & Wolf-Watz, M. (2018). Molecular mechanism of ATP versus GTP selectivity of adenylate kinase. Proceedings of the National Academy of Sciences of the United States of America, 115(12), 3012-3017
Open this publication in new window or tab >>Molecular mechanism of ATP versus GTP selectivity of adenylate kinase
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2018 (English)In: Proceedings of the National Academy of Sciences of the United States of America, ISSN 0027-8424, E-ISSN 1091-6490, Vol. 115, no 12, p. 3012-3017Article in journal (Refereed) Published
Abstract [en]

Enzymatic substrate selectivity is critical for the precise control of metabolic pathways. In cases where chemically related substrates are present inside cells, robust mechanisms of substrate selectivity are required. Here, we report the mechanism utilized for catalytic ATP versus GTP selectivity during adenylate kinase (Adk) -mediated phosphorylation of AMP. Using NMR spectroscopy we found that while Adk adopts a catalytically competent and closed structural state in complex with ATP, the enzyme is arrested in a catalytically inhibited and open state in complex with GTP. X-ray crystallography experiments revealed that the interaction interfaces supporting ATP and GTP recognition, in part, are mediated by coinciding residues. The mechanism provides an atomic view on how the cellular GTP pool is protected from Adk turnover, which is important because GTP has many specialized cellular functions. In further support of this mechanism, a structure-function analysis enabled by synthesis of ATP analogs suggests that a hydrogen bond between the adenine moiety and the backbone of the enzyme is vital for ATP selectivity. The importance of the hydrogen bond for substrate selectivity is likely general given the conservation of its location and orientation across the family of eukaryotic protein kinases.

Keywords
adenylate kinase, selectivity, ATP, GTP, mechanism
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:umu:diva-145883 (URN)10.1073/pnas.1721508115 (DOI)000427829500063 ()29507216 (PubMedID)
Available from: 2018-03-20 Created: 2018-03-20 Last updated: 2018-06-09Bibliographically approved
Shukla, L., Moodie, L. W. K., Kindahl, T. & Hedberg, C. (2018). Synthesis and Spectroscopic Properties of Fluorinated Coumarin Lysine Derivatives. Journal of Organic Chemistry, 83(8), 4792-4799
Open this publication in new window or tab >>Synthesis and Spectroscopic Properties of Fluorinated Coumarin Lysine Derivatives
2018 (English)In: Journal of Organic Chemistry, ISSN 0022-3263, E-ISSN 1520-6904, Vol. 83, no 8, p. 4792-4799Article in journal (Refereed) Published
Abstract [en]

The site-selective incorporation of fluorescent amino acids into proteins has emerged as a valuable alternative to expressible protein reporters. For successful application, a robust and scalable, yet flexible, route to non-natural amino acids is required. This work describes an improved synthesis of coumarin-conjugated lysine derivatives where fluorinated variants are accessed. These analogues can be utilized at low pH and should find application probing biological processes that operate under acidic conditions.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2018
National Category
Organic Chemistry
Identifiers
urn:nbn:se:umu:diva-148028 (URN)10.1021/acs.joc.7b03214 (DOI)000430898500046 ()29595263 (PubMedID)
Available from: 2018-05-30 Created: 2018-05-30 Last updated: 2018-06-09Bibliographically approved
Moodie, L. W. K., Chammaa, S., Kindahl, T. & Hedberg, C. (2017). Palladium-Mediated Approach to Coumarin-Functionalized Amino Acids. Organic Letters, 19(11), 2797-2800
Open this publication in new window or tab >>Palladium-Mediated Approach to Coumarin-Functionalized Amino Acids
2017 (English)In: Organic Letters, ISSN 1523-7060, E-ISSN 1523-7052, Vol. 19, no 11, p. 2797-2800Article in journal (Refereed) Published
Abstract [en]

Incorporation of the fluorogenic l-(7-hydroxycoumarin-4-yl)ethylglycine into proteins is a valuable biological tool. Coumarins are typically accessed via the Pechmann reaction, which requires acidic conditions and lacks substrate flexibility. A Pd-mediated coupling is described between o-methoxyboronic acids and a glutamic acid derived (Z)-vinyl triflate, forming latent coumarins. Global deprotection with BBr3 forms the coumarin scaffold in a single step. This mild and scalable route yielded five analogues, including a probe suitable for use at lower pH.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2017
National Category
Organic Chemistry
Identifiers
urn:nbn:se:umu:diva-137386 (URN)10.1021/acs.orglett.7b00854 (DOI)000402850900006 ()28497693 (PubMedID)
Available from: 2017-07-06 Created: 2017-07-06 Last updated: 2018-06-09Bibliographically approved
Martinez, N. E., Zimmermann, T. J., Goosmann, C., Alexander, T., Hedberg, C., Ziegler, S., . . . Waldmann, H. (2017). Tetrahydroisoquinolines: New Inhibitors of Neutrophil Extracellular Trap (NET) Formation. ChemBioChem (Print), 18(10), 888-893
Open this publication in new window or tab >>Tetrahydroisoquinolines: New Inhibitors of Neutrophil Extracellular Trap (NET) Formation
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2017 (English)In: ChemBioChem (Print), ISSN 1439-4227, E-ISSN 1439-7633, Vol. 18, no 10, p. 888-893Article in journal (Refereed) Published
Abstract [en]

Neutrophils are short-lived leukocytes that migrate to sites of infection as part of the acute immune response, where they phagocytose, degranulate, and form neutrophil extracellular traps (NETs). During NET formation, the nuclear lobules of neutrophils disappear and the chromatin expands and, accessorized with neutrophilic granule proteins, is expelled. NETs can be pathogenic in, for example, sepsis, cancer, and autoimmune and cardiovascular diseases. Therefore, the identification of inhibitors of NET formation is of great interest. Screening of a focused library of natural-product-inspired compounds by using a previously validated phenotypic NET assay identified a group of tetrahydroisoquinolines as new NET formation inhibitors. This compound class opens up new avenues for the study of cellular death through NET formation (NETosis) at different stages, and might inspire new medicinal chemistry programs aimed at NET-dependent diseases.

Place, publisher, year, edition, pages
Wiley-VCH Verlagsgesellschaft, 2017
Keywords
inhibitors, NETosis, neutrophil extracellular traps (NETs), structure-activity relationships, trahydroisoquinolines
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:umu:diva-135969 (URN)10.1002/cbic.201600650 (DOI)000401868700004 ()28240414 (PubMedID)
Available from: 2017-06-27 Created: 2017-06-27 Last updated: 2018-06-09Bibliographically approved
Qian, Y., Schuermann, M., Janning, P., Hedberg, C. & Waldmann, H. (2016). Activity-Based Proteome Profiling Probes Based on Woodward's Reagent K with Distinct Target Selectivity. Angewandte Chemie International Edition, 55(27), 7766-7771
Open this publication in new window or tab >>Activity-Based Proteome Profiling Probes Based on Woodward's Reagent K with Distinct Target Selectivity
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2016 (English)In: Angewandte Chemie International Edition, ISSN 1433-7851, E-ISSN 1521-3773, Vol. 55, no 27, p. 7766-7771Article in journal (Refereed) Published
Abstract [en]

Woodward's reagent K (WRK) is a reactive heterocyclic compound that has been employed in protein chemistry to covalently and unspecifically label proteins at nucleophilic amino acids, notably at histidine and cysteine. We have developed a panel of WRK-derived activity-based probes and show that surprisingly and unexpectedly, these probes are fairly selective for a few proteins in the human proteome. The WRK-derived probes show unique reactivity towards the catalytic N-terminal proline in the macrophage migration inhibitory factor (MIF) and can be used to label and, if equipped with a fluorophore, to image MIF activities in living cells.

National Category
Analytical Chemistry
Identifiers
urn:nbn:se:umu:diva-126530 (URN)10.1002/anie.201602666 (DOI)000383252900030 ()27159346 (PubMedID)
Available from: 2016-10-26 Created: 2016-10-10 Last updated: 2018-06-09Bibliographically approved
Schell, U., Simon, S., Sahr, T., Hager, D., Albers, M. F., Kessler, A., . . . Hilbi, H. (2016). The α-hydroxyketone LAI-1 regulates motility, Lqs-dependent phosphorylation signaling and gene expression of Legionella pneumophila. Molecular Microbiology, 99(4), 778-793
Open this publication in new window or tab >>The α-hydroxyketone LAI-1 regulates motility, Lqs-dependent phosphorylation signaling and gene expression of Legionella pneumophila
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2016 (English)In: Molecular Microbiology, ISSN 0950-382X, E-ISSN 1365-2958, Vol. 99, no 4, p. 778-793Article in journal (Refereed) Published
Abstract [en]

The causative agent of Legionnaires' disease, Legionella pneumophila, employs the autoinducer compound LAI-1 (3-hydroxypentadecane-4-one) for cell–cell communication. LAI-1 is produced and detected by the Lqs (Legionella quorum sensing) system, comprising the autoinducer synthase LqsA, the sensor kinases LqsS and LqsT, as well as the response regulator LqsR. Lqs-regulated processes include pathogen–host interactions, production of extracellular filaments and natural competence for DNA uptake. Here we show that synthetic LAI-1 promotes the motility of L. pneumophila by signalling through LqsS/LqsT and LqsR. Upon addition of LAI-1, autophosphorylation of LqsS/LqsT by [γ-32P]-ATP was inhibited in a dose-dependent manner. In contrast, the Vibrio cholerae autoinducer CAI-1 (3-hydroxytridecane-4-one) promoted the phosphorylation of LqsS (but not LqsT). LAI-1 did neither affect the stability of phospho-LqsS or phospho-LqsT, nor the dephosphorylation by LqsR. Transcriptome analysis of L. pneumophila treated with LAI-1 revealed that the compound positively regulates a number of genes, including the non-coding RNAs rsmY and rsmZ, and negatively regulates the RNA-binding global regulator crsA. Accordingly, LAI-1 controls the switch from the replicative to the transmissive growth phase of L. pneumophila. In summary, the findings indicate that LAI-1 regulates motility and the biphasic life style of L. pneumophila through LqsS- and LqsT-dependent phosphorylation signalling.

Place, publisher, year, edition, pages
Wiley-Blackwell, 2016
National Category
Microbiology
Identifiers
urn:nbn:se:umu:diva-114331 (URN)10.1111/mmi.13265 (DOI)000370338900012 ()26538361 (PubMedID)
Note

Article first published online: 27 NOV 2015

Available from: 2016-01-18 Created: 2016-01-18 Last updated: 2018-06-07Bibliographically approved
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