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Jacobsson, Erik
Publications (10 of 13) Show all publications
Jacobsson, E. (2019). Studies on cysteine-rich peptides from Nemertea and Violaceae: Proteomic and transcriptomic discovery and characterization. (Doctoral dissertation). Uppsala: Acta Universitatis Upsaliensis
Open this publication in new window or tab >>Studies on cysteine-rich peptides from Nemertea and Violaceae: Proteomic and transcriptomic discovery and characterization
2019 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The overall aims of the projects included in this thesis were to discover, synthesize and characterize disulphide-stabilized peptides from marine worms (Nemertea sp.) and plants (Viola sp.). 

One of the main outcomes of this thesis is the discovery of a new family of highly active cysteine-rich toxins, alpha nemertides, from nemertean worms (paper II). Functional characterization and production routes of nemertides were further explored (papers II-III). In addition, 12 new cyclotides from the bog violet were discovered (paper I). Finally, transcriptomes and mucus of the Antarctic nemertean Parborlasia corrugatus were investigated for toxin content (paper IV).

 In paper I wild-type leaf and callus tissue of the endangered bog violet, V. uliginosa, were analyzed using transcriptomics and LC-MS, resulting in the discovery of 12 new cyclotides (i.e. cysteine-rich cyclic peptides). In addition, cyclotide expression under different cell-growth conditions was monitored.

In paper II  the discovery and initial characterization of a new family of highly active peptides, the alpha nemertides, from the epidermal mucus of the world’s longest animal; Lineus longissimus is described. The most abundant alpha nemertide, alpha-1, was extracted in minute amounts, prompting the use solid phase peptide synthesis (SPPS) for further characterization. The tertiary structure of alpha-1 was elucidated and revealed an inhibitory cystine knot (ICK) framework. The knotted core-structure is similar to the cyclic cystine knot (CCK) motif, found in the cyclotides described in paper I.

In manuscript III, the production route established in paper II was used to produce nemertides alpha 1-7. These were tested in vivo in an Artemia microwell assay as well as on an extended panel of voltage-gated sodium channels (NaV1.1 – 1.8 and BgNaV1). All seven alpha nemertides were highly active in the in vivo Artemia assay with EC50 values in the sub to low µM range. The alpha nemertides were also active in the NaVs tested. However, differences in the activity profiles were observed, indicating an opportunity for future optimization of alpha nemertides to reach higher specificity to certain NaV subtypes.

In manuscript IV, the exploration of nemertide toxins was extended to include the Antarctic P. corrugatus. Resulting findings include a set of cysteine-rich peptides, some similar to the nemertides previously discovered in paper II. Two purified peptides and one fraction were evaluated for their membranolytic activity.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2019. p. 66
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Pharmacy, ISSN 1651-6192 ; 277
Keywords
Peptide toxin, cystine knot, nemertide, cyclotide, nemertea.
National Category
Pharmacology and Toxicology
Research subject
Pharmacognosy
Identifiers
urn:nbn:se:uu:diva-390885 (URN)978-91-513-0719-0 (ISBN)
Public defence
2019-09-20, BMC A1:107a, Husargatan 3, Uppsala, 09:15 (English)
Opponent
Supervisors
Available from: 2019-08-30 Created: 2019-08-15 Last updated: 2019-09-17
Göransson, U., Jacobsson, E., Strand, M. & Andersson, H. S. (2019). The Toxins of Nemertean Worms. Toxins, 11(2), Article ID 120.
Open this publication in new window or tab >>The Toxins of Nemertean Worms
2019 (English)In: Toxins, ISSN 2072-6651, E-ISSN 2072-6651, Vol. 11, no 2, article id 120Article, review/survey (Refereed) Published
Abstract [en]

Most ribbon worms (phylum: Nemertea) are found in marine environments, where they act as predators and scavengers. They are characterized by an eversible proboscis that is used to hunt for prey and thick mucus covering their skin. Both proboscis and epidermal mucus mediate toxicity to predators and preys. Research into the chemical nature of the substances that render toxicity has not been extensive, but it has nevertheless led to the identification of several compounds of potential medicinal use or for application in biotechnology. This review provides a complete account of the current status of research into nemertean toxins.

Place, publisher, year, edition, pages
MDPI, 2019
Keywords
Anabaseine, cytotoxin, DMXBA, nemertea, nemertide, parborlysin, ribbon worm, tetrodotoxin
National Category
Zoology
Identifiers
urn:nbn:se:uu:diva-380672 (URN)10.3390/toxins11020120 (DOI)000460801500062 ()30781381 (PubMedID)
Funder
Swedish Research Council Formas, 2018-00613Swedish Research Council, 214-3327; 2018-005403The Crafoord Foundation, 20160810
Available from: 2019-04-01 Created: 2019-04-01 Last updated: 2019-04-01Bibliographically approved
Jacobsson, E., Andersson, H. S., Strand, M., Peigneur, S., Eriksson, C., Lodén, H., . . . Göransson, U. (2018). Peptide ion channel toxins from the bootlace worm, the longest animal on Earth. Scientific Reports, 8, Article ID 4596.
Open this publication in new window or tab >>Peptide ion channel toxins from the bootlace worm, the longest animal on Earth
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2018 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 8, article id 4596Article in journal (Refereed) Published
Abstract [en]

Polypeptides from animal venoms have found important uses as drugs, pharmacological tools, and within biotechnological and agricultural applications. We here report a novel family of cystine knot peptides from nemertean worms, with potent activity on voltage-gated sodium channels. These toxins, named the alpha-nemertides, were discovered in the epidermal mucus of Lineus longissimus, the 'bootlace worm' known as the longest animal on earth. The most abundant peptide, the 31-residue long alpha-1, was isolated, synthesized, and its 3D NMR structure determined. Transcriptome analysis including 17 species revealed eight alpha-nemertides, mainly distributed in the genus Lineus. alpha-1 caused paralysis and death in green crabs (Carcinus maenas) at 1 mu g/kg (similar to 300 pmol/kg). It showed profound effect on invertebrate voltage-gated sodium channels (e.g. Blattella germanica Na(v)1) at low nanomolar concentrations. Strong selectivity for insect over human sodium channels indicates that a-nemertides can be promising candidates for development of bioinsecticidal agents.

Place, publisher, year, edition, pages
NATURE PUBLISHING GROUP, 2018
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:uu:diva-351585 (URN)10.1038/s41598-018-22305-w (DOI)000428029600001 ()29567943 (PubMedID)
Funder
Swedish Research Council, 2014-3327]
Available from: 2018-05-29 Created: 2018-05-29 Last updated: 2019-08-15Bibliographically approved
Park, S., Yoo, K.-O., Marcussen, T., Backlund, A., Jacobsson, E., Rosengren, K. J., . . . Göransson, U. (2017). Cyclotide Evolution: Insights from the Analyses of Their Precursor Sequences, Structures and Distribution in Violets (Viola). Frontiers in Plant Science, 8, Article ID 2058.
Open this publication in new window or tab >>Cyclotide Evolution: Insights from the Analyses of Their Precursor Sequences, Structures and Distribution in Violets (Viola)
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2017 (English)In: Frontiers in Plant Science, ISSN 1664-462X, E-ISSN 1664-462X, Vol. 8, article id 2058Article in journal (Refereed) Published
Abstract [en]

Cyclotides are a family of plant proteins that are characterized by a cyclic backbone and a knotted disulfide topology. Their cyclic cystine knot (CCK) motif makes them exceptionally resistant to thermal, chemical, and enzymatic degradation. By disrupting cell membranes, the cyclotides function as host defense peptides by exhibiting insecticidal, anthelmintic, antifouling, and molluscicidal activities. In this work, we provide the first insight into the evolution of this family of plant proteins by studying the Violaceae, in particular species of the genus Viola. We discovered 157 novel precursor sequences by the transcriptomic analysis of six Viola species: V. albida var. takahashii, V. mandshurica, V. orientalis, V. verecunda, V. acuminata, and V. canadensis. By combining these precursor sequences with the phylogenetic classification of Viola, we infer the distribution of cyclotides across 63% of the species in the genus (i.e., ~380 species). Using full precursor sequences from transcriptomes, we show an evolutionary link to the structural diversity of the cyclotides, and further classify the cyclotides by sequence signatures from the non-cyclotide domain. Also, transcriptomes were compared to cyclotide expression on a peptide level determined using liquid chromatography-mass spectrometry. Furthermore, the novel cyclotides discovered were associated with the emergence of new biological functions.

Keywords
cyclotide evolution, viola phylogeny, sequence signature, cyclotide precursor, neofunctionality, novel cyclotide, precursor domain
National Category
Plant Biotechnology
Identifiers
urn:nbn:se:uu:diva-339782 (URN)10.3389/fpls.2017.02058 (DOI)000418117800001 ()29326730 (PubMedID)
Funder
Swedish Research Council, 2012-5063
Available from: 2018-02-09 Created: 2018-02-09 Last updated: 2018-02-09Bibliographically approved
Göransson, U., Gunasekera, S., Malik, S., Park, S., Slazak, B., Jacobsson, E., . . . Strömstedt, A. A. (2016). Peptide biodiscovery from plants and animals: structure to function. Paper presented at 9th Joint Meeting of AFERP, ASP, GA, JSP, PSE and SIF, JUL 24-27, 2016, Copenhagen, DENMARK. Planta Medica, 82
Open this publication in new window or tab >>Peptide biodiscovery from plants and animals: structure to function
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2016 (English)In: Planta Medica, ISSN 0032-0943, E-ISSN 1439-0221, Vol. 82Article in journal, Meeting abstract (Other academic) Published
Keywords
Peptide biodiscovery, cyclotides, sponge peptides, cacti
National Category
Basic Medicine
Identifiers
urn:nbn:se:uu:diva-346852 (URN)10.1055/s-0036-1596156 (DOI)000411789300953 ()
Conference
9th Joint Meeting of AFERP, ASP, GA, JSP, PSE and SIF, JUL 24-27, 2016, Copenhagen, DENMARK
Available from: 2018-03-27 Created: 2018-03-27 Last updated: 2018-03-27Bibliographically approved
Jacobsson, E., Andersson, H. S., Strand, M., Lebbe, E., Eriksson, C., Peigneur, S., . . . Göransson, U. (2016). Peptide toxins from the longest animal on earth. Paper presented at 9th Joint Meeting of AFERP, ASP, GA, JSP, PSE and SIF, JUL 24-27, 2016, Copenhagen, DENMARK. Planta Medica, 82
Open this publication in new window or tab >>Peptide toxins from the longest animal on earth
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2016 (English)In: Planta Medica, ISSN 0032-0943, E-ISSN 1439-0221, Vol. 82Article in journal, Meeting abstract (Other academic) Published
Keywords
peptide toxin, nemertea, neurotoxin
National Category
Pharmacology and Toxicology
Identifiers
urn:nbn:se:uu:diva-346855 (URN)10.1055/s-0036-1596160 (DOI)000411789300986 ()
Conference
9th Joint Meeting of AFERP, ASP, GA, JSP, PSE and SIF, JUL 24-27, 2016, Copenhagen, DENMARK
Available from: 2018-03-26 Created: 2018-03-26 Last updated: 2018-03-26Bibliographically approved
Strand, M., Hedström, M., Seth, H., McEvoy, E. G., Jacobsson, E., Göransson, U., . . . Sundberg, P. (2016). The Bacterial (Vibrio alginolyticus) Production of Tetrodotoxin in the Ribbon Worm Lineus longissimus-Just a False Positive?. Marine Drugs, 14(4), Article ID 63.
Open this publication in new window or tab >>The Bacterial (Vibrio alginolyticus) Production of Tetrodotoxin in the Ribbon Worm Lineus longissimus-Just a False Positive?
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2016 (English)In: Marine Drugs, ISSN 1660-3397, E-ISSN 1660-3397, Vol. 14, no 4, article id 63Article in journal (Refereed) Published
Abstract [en]

We test previous claims that the bacteria Vibrio alginolyticus produces tetrodotoxin (TTX) when living in symbiosis with the nemertean Lineus longissimus by a setup with bacteria cultivation for TTX production. Toxicity experiments on the shore crab, Carcinus maenas, demonstrated the presence of a paralytic toxin, but evidence from LC-MS and electrophysiological measurements of voltage-gated sodium channel-dependent nerve conductance in male Wistar rat tissue showed conclusively that this effect did not originate from TTX. However, a compound of similar molecular weight was found, albeit apparently non-toxic, and with different LC retention time and MS/MS fragmentation pattern than those of TTX. We conclude that C. maenas paralysis and death likely emanate from a compound <5 kDa, and via a different mechanism of action than that of TTX. The similarity in mass between TTX and the Vibrio-produced low-molecular-weight, non-toxic compound invokes that thorough analysis is required when assessing TTX production. Based on our findings, we suggest that re-examination of some published claims of TTX production may be warranted.

Keywords
LC-MS; mucus; axonal conductance; Vibrio; nemertean; tetrodotoxin
National Category
Pharmaceutical Sciences
Identifiers
urn:nbn:se:uu:diva-289125 (URN)10.3390/md14040063 (DOI)000374588000002 ()27023570 (PubMedID)
Available from: 2016-04-28 Created: 2016-04-28 Last updated: 2018-01-10Bibliographically approved
Andersson, H. S., Jacobsson, E., Eriksson, C., Hedstrom, M., Seth, H., Sundberg, P., . . . Göransson, U. (2016). The toxicity of ribbon worms: Alpha-nemertides or tetrodotoxin, or both?. Paper presented at 9th Joint Meeting of AFERP, ASP, GA, JSP, PSE and SIF, JUL 24-27, 2016, Copenhagen, DENMARK. Planta Medica, 82
Open this publication in new window or tab >>The toxicity of ribbon worms: Alpha-nemertides or tetrodotoxin, or both?
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2016 (English)In: Planta Medica, ISSN 0032-0943, E-ISSN 1439-0221, Vol. 82Article in journal, Meeting abstract (Other academic) Published
National Category
Pharmacology and Toxicology
Identifiers
urn:nbn:se:uu:diva-346848 (URN)10.1055/s-0036-1596617 (DOI)000411789300441 ()
Conference
9th Joint Meeting of AFERP, ASP, GA, JSP, PSE and SIF, JUL 24-27, 2016, Copenhagen, DENMARK
Available from: 2018-03-27 Created: 2018-03-27 Last updated: 2018-03-27Bibliographically approved
Slazak, B., Jacobsson, E., Kuta, E. & Goransson, U. (2015). Exogenous plant hormones and cyclotide expression in Viola uliginosa (Violaceae). Phytochemistry, 117, 527-536
Open this publication in new window or tab >>Exogenous plant hormones and cyclotide expression in Viola uliginosa (Violaceae)
2015 (English)In: Phytochemistry, ISSN 0031-9422, E-ISSN 1873-3700, Vol. 117, p. 527-536Article in journal (Refereed) Published
Abstract [en]

Plants from Violaceae produce cyclotides, peptides characterized by a circular peptide backbone and a cystine knot. This signature motif gives stability that can harness a wide spectrum of biological activities, with implications in plant defense and with applications in medicine and biotechnology. In the current work, cyclotide expressing in vitro cultures were established from Viola uliginosa. These cultures are useful models for studying biosynthesis of cyclotides and can also be used in their production. The cyclotide expression pattern is shown to be dependent on exogenous plant growth regulators, both on peptide and gene expression levels. The highest yields of cyclotides were obtained on media containing only a cytokinin and were correlated with storage material accumulation. Exposure to auxins decreased cyclotide production and caused shifting of the biosynthesis pattern to root specific cyclotides. The response to stimuli in terms of cyclotide expression pattern appears to be developmental, and related to polar auxin transportation and the auxin/cytokinin ratio regulating tissue differentiation. By the use of whole transcriptome shotgun sequencing (WTSS) and peptidomics, 20 cyclotide sequences from V. uliginosa (including 12 new) and 12 complete precursor proteins could be identified. The most abundant cyclotides were cycloviolacin O3 (CyO3), CyO8 and CyO13. A suspension culture was obtained that grew exponentially with a doubling time of approximately 3 days. After ten days of growth, the culture provided a yield of more than 4 mg CyO13 per gram dry mass.

Keywords
Viola uliginosa (Violaceae), Cyclotides, In vitro culture, Plant growth regulators, Whole transcriptome shotgun sequencing, Mass spectrometry
National Category
Botany Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:uu:diva-264668 (URN)10.1016/j.phytochem.2015.07.016 (DOI)000361253300055 ()26246035 (PubMedID)
Funder
Swedish Research Council, 621-2007-5167Swedish Foundation for Strategic Research , F06-0058
Available from: 2015-10-16 Created: 2015-10-15 Last updated: 2019-08-15
Hussein, J., Chi, C. N., Tibell Savić, S., Tibuhwa, D., Jacobsson, E., Rosengren, J., . . . Göransson, U.Cysteine-rich peptide from the gigantic edible mushroom Kusaghiporia usambarensis (Laetiporaceae).
Open this publication in new window or tab >>Cysteine-rich peptide from the gigantic edible mushroom Kusaghiporia usambarensis (Laetiporaceae)
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(English)Manuscript (preprint) (Other academic)
Abstract [en]

Cysteine-rich peptides are produced by various organisms across all kingdoms and have triggered an interest in isolation of molecules for novel drug development. In this study, we report a novel cysteine-rich peptide, kusaghitide, isolated from the gigantic medicinal mushroom Kusaghiporia usambarensis. It is highly expressed in the K. usambarensis transcriptome and it is the most abundant compound in the methanol-water extract. The 54 amino acid residue long peptide was isolated through aqueous methanol 50% and a sample was reduced, alkylated and cleaved enzymatically. De novo sequencing was done by LC-MS/MS and obtained sequences were used for mining the transcriptome to search for the complete gene. The peptide was recombinantly expressed in One Shot BL21 Star Escherichia coli using lysogenic broth and minimal media. Its 3D NMR structure was determined using 2D and 3D NMR. Three hypothetical protein sequences similar to kusaghitide originate from Laetiporus sulphureusWolfiporia cocos and Sparassis crispa with per cent similarity of 76% and 58% and 53% respectively and were found by BLAST search in the NCBI database. Kusaghitide did not inhibit the growth of either Escherichia coli or Staphylococcus aureus. This is first report of a peptide from K. usambarensis in Laetiporaceae.

Keywords
Kusaghiporia, medicinal mushroom, recombinant protein production, cysteine-rich peptides, peptide structure.
National Category
Natural Sciences
Identifiers
urn:nbn:se:uu:diva-405387 (URN)
Available from: 2020-02-28 Created: 2020-02-28 Last updated: 2020-02-28
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