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Persson, Karina
Publications (10 of 26) Show all publications
Persson, K. & Backman, L. (2018). Crystallization of recombinant α-actinin and related proteins. In: Teresa L. Singleton (Ed.), Schizosaccharomyces pombe: methods and protocols (pp. 95-103). Humana Press
Open this publication in new window or tab >>Crystallization of recombinant α-actinin and related proteins
2018 (English)In: Schizosaccharomyces pombe: methods and protocols / [ed] Teresa L. Singleton, Humana Press, 2018, p. 95-103Chapter in book (Other academic)
Abstract [en]

When it comes to crystallization each protein is unique. It can never be predicted beforehand in which condition the particular protein will crystallize or even if it is possible to crystallize. Still, by following some simple checkpoints the chances of obtaining crystals are increased. The primary checkpoints are purity, stability, concentration, and homogeneity. High-quality protein crystals are needed. This protocol will allow an investigator to: clone, express, and crystallize a protein of interest.

Place, publisher, year, edition, pages
Humana Press, 2018
Series
Methods in Molecular Biology, ISSN 1064-3745, E-ISSN 1940-6029 ; 1721
Keywords
Protein purification, Crystallization, Optimization
National Category
Biochemistry and Molecular Biology
Research subject
Biochemistry
Identifiers
urn:nbn:se:umu:diva-145559 (URN)10.1007/978-1-4939-7546-4_9 (DOI)978-1-4939-7545-7 (ISBN)978-1-4939-7546-4 (ISBN)
Available from: 2018-03-09 Created: 2018-03-09 Last updated: 2019-03-14Bibliographically approved
Dongre, M., Singh, B., Aung, K. M., Larsson, P., Miftakhova, R. R., Persson, K., . . . Wai, S. N. (2018). Flagella-mediated secretion of a novel Vibrio cholerae cytotoxin affecting both vertebrate and invertebrate hosts. Communications Biology, 1, Article ID 59.
Open this publication in new window or tab >>Flagella-mediated secretion of a novel Vibrio cholerae cytotoxin affecting both vertebrate and invertebrate hosts
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2018 (English)In: Communications Biology, ISSN 2399-3642, Vol. 1, article id 59Article in journal (Refereed) Published
Abstract [en]

Using Caenorhabditis elegans as an infection host model for Vibrio cholerae predator interactions, we discovered a bacterial cytotoxin, MakA, whose function as a virulence factor relies on secretion via the flagellum channel in a proton motive force-dependent manner. The MakA protein is expressed from the polycistronic makDCBA (motility-associated killing factor) operon. Bacteria expressing makDCBA induced dramatic changes in intestinal morphology leading to a defecation defect, starvation and death in C. elegans. The Mak proteins also promoted V. cholerae colonization of the zebrafish gut causing lethal infection. A structural model of purified MakA at 1.9 Å resolution indicated similarities to members of a superfamily of bacterial toxins with unknown biological roles. Our findings reveal an unrecognized role for V. cholerae flagella in cytotoxin export that may contribute both to environmental spread of the bacteria by promoting survival and proliferation in encounters with predators, and to pathophysiological effects during infections.

Place, publisher, year, edition, pages
Springer Nature Publishing AG, 2018
National Category
Microbiology in the medical area
Research subject
Infectious Diseases; Molecular Biology
Identifiers
urn:nbn:se:umu:diva-155563 (URN)10.1038/s42003-018-0065-z (DOI)000461126500059 ()30271941 (PubMedID)
Available from: 2019-01-22 Created: 2019-01-22 Last updated: 2019-04-04Bibliographically approved
Hall, M., Hasegawa, Y., Yoshimura, F. & Persson, K. (2018). Structural and functional characterization of shaft, anchor, and tip proteins of the Mfa1 fimbria from the periodontal pathogen Porphyromonas gingivalis. Scientific Reports, 8, Article ID 1793.
Open this publication in new window or tab >>Structural and functional characterization of shaft, anchor, and tip proteins of the Mfa1 fimbria from the periodontal pathogen Porphyromonas gingivalis
2018 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 8, article id 1793Article in journal (Refereed) Published
Abstract [en]

Very little is known about how fimbriae of Bacteroidetes bacteria are assembled. To shed more light on this process, we solved the crystal structures of the shaft protein Mfa1, the regulatory protein Mfa2, and the tip protein Mfa3 from the periodontal pathogen Porphyromonas gingivalis. Together these build up part of the Mfa1 fimbria and represent three of the five proteins, Mfa1-5, encoded by the mfa1 gene cluster. Mfa1, Mfa2 and Mfa3 have the same overall fold i.e., two β-sandwich domains. Upon polymerization, the first β-strand of the shaft or tip protein is removed by indigenous proteases. Although the resulting void is expected to be filled by a donor-strand from another fimbrial protein, the mechanism by which it does so is still not established. In contrast, the first β-strand in Mfa2, the anchoring protein, is firmly attached by a disulphide bond and is not cleaved. Based on the structural information, we created multiple mutations in P. gingivalis and analysed their effect on fimbrial polymerization and assembly in vivo. Collectively, these data suggest an important role for the C-terminal tail of Mfa1, but not of Mfa3, affecting both polymerization and maturation of downstream fimbrial proteins.

National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:umu:diva-144501 (URN)10.1038/s41598-018-20067-z (DOI)000423430400005 ()29379120 (PubMedID)
Available from: 2018-02-05 Created: 2018-02-05 Last updated: 2018-06-09Bibliographically approved
Persson, K., Hall, M., Heidler, T. & Hasegawa, Y. (2018). Structural studies of the five pilin proteins building up the type-V pilus Mfa1 of Porphyromonas gingivalis. Acta Crystallographica - Section A : Foundations and Advances, 74, E425-E425
Open this publication in new window or tab >>Structural studies of the five pilin proteins building up the type-V pilus Mfa1 of Porphyromonas gingivalis
2018 (English)In: Acta Crystallographica - Section A : Foundations and Advances, ISSN 2053-2733, Vol. 74, p. E425-E425Article in journal, Meeting abstract (Other academic) Published
Place, publisher, year, edition, pages
International Union of Crystallography, 2018
Keywords
pill, polynraerizaticxaa, bacteria
Identifiers
urn:nbn:se:umu:diva-161753 (URN)10.1107/S2053273318088800 (DOI)000474406600648 ()
Available from: 2019-07-25 Created: 2019-07-25 Last updated: 2019-07-25Bibliographically approved
Backman, L. & Persson, K. (2018). The no-nonsens SDS-PAGE. In: Teresa L. Singleton (Ed.), Schizosaccharomyces pombe: methods and protocols (pp. 89-94). Humana Press
Open this publication in new window or tab >>The no-nonsens SDS-PAGE
2018 (English)In: Schizosaccharomyces pombe: methods and protocols / [ed] Teresa L. Singleton, Humana Press, 2018, p. 89-94Chapter in book (Other academic)
Abstract [en]

The discontinuous polyacrylamide gel electrophoresis system devised by Laemmli (Nature 227:680–685, 1970) has not only been used in numerous laboratories but has also been modified in several ways since its birth. In our laboratories, we use a modified Laemmli SDS-PAGE system for following protein purification as well as for analysis of certain protein-protein interactions, mainly involving filametous actin.

Place, publisher, year, edition, pages
Humana Press, 2018
Series
Methods in molecular biology, ISSN 1064-3745, E-ISSN 1940-6029 ; 1721
Keywords
SDS-PAGE, polyacrylamide gel electrophoresis
National Category
Biochemistry and Molecular Biology
Research subject
Biochemistry
Identifiers
urn:nbn:se:umu:diva-145557 (URN)10.1007/978-1-4939-7546-4_8 (DOI)978-1-4939-7545-7 (ISBN)978-1-4939-7546-4 (ISBN)
Available from: 2018-03-09 Created: 2018-03-09 Last updated: 2019-03-14Bibliographically approved
Esberg, A., Sheng, N., Mårell, L., Claesson, R., Persson, K., Borén, T. & Strömberg, N. (2017). Streptococcus Mutans Adhesin Biotypes that Match and Predict Individual Caries Development. EBioMedicine, 24, 205-215
Open this publication in new window or tab >>Streptococcus Mutans Adhesin Biotypes that Match and Predict Individual Caries Development
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2017 (English)In: EBioMedicine, ISSN 0360-0637, E-ISSN 2352-3964, Vol. 24, p. 205-215Article in journal (Refereed) Published
Abstract [en]

Dental caries, which affects billions of people, is a chronic infectious disease that involves Streptococcus mutans, which is nevertheless a poor predictor of individual caries development. We therefore investigated if adhesin types of S.mutans with sucrose-independent adhesion to host DMBT1 (i.e. SpaP A, B or C) and collagen (i.e. Cnm, Cbm) match and predict individual differences in caries development. The adhesin types were measured in whole saliva by qPCR in 452 12-year-old Swedish children and related to caries at baseline and prospectively at a 5-year follow-up. Strains isolated from the children were explored for genetic and phenotypic properties. The presence of SpaP B and Cnm subtypes coincided with increased 5-year caries increment, and their binding to DMBT1 and saliva correlated with individual caries scores. The SpaP B subtypes are enriched in amino acid substitutions that coincided with caries and binding and specify biotypes of S. mutans with increased acid tolerance. The findings reveal adhesin subtypes of S. mutans that match and predict individual differences in caries development and provide a rationale for individualized oral care.

Keywords
Adhesion, Chronic infections, Dental caries, SpaP, Streptococcus mutans, Virulence
National Category
Dentistry
Identifiers
urn:nbn:se:umu:diva-140203 (URN)10.1016/j.ebiom.2017.09.027 (DOI)000414392900033 ()28958656 (PubMedID)
Funder
Swedish Research Council, 10906
Available from: 2017-10-03 Created: 2017-10-03 Last updated: 2018-06-09Bibliographically approved
Hall, M., Wagner, R., Tam, L. X., Funk, C. & Persson, K. (2017). The HhoA protease from Synechocystis sp. PCC 6803 – Novel insights into structure and activity regulation. Journal of Structural Biology, 198(3), 147-153
Open this publication in new window or tab >>The HhoA protease from Synechocystis sp. PCC 6803 – Novel insights into structure and activity regulation
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2017 (English)In: Journal of Structural Biology, ISSN 1047-8477, E-ISSN 1095-8657, Vol. 198, no 3, p. 147-153Article in journal (Refereed) Published
Abstract [en]

Abstract Proteases play a vital role in the removal of proteins, which become damaged due to temperature or oxidative stress. Important to this process in the cyanobacterium Synechocystis sp. PCC6803 is the family of Deg/HtrA proteases; HhoA (sll1679), HhoB (sll1427) and HtrA (slr1204). While previous studies have elucidated the structures of Deg/HtrA proteases from Escherichia coli and from the chloroplast of the higher plant Arabidopsis thaliana, no structural data have been available for any Deg/HtrA protease from cyanobacteria, the evolutionary ancestor of the chloroplast. To gain a deeper insight into the molecular mechanisms and regulation of these proteins we have solved the structure of the Synechocystis HhoA protease in complex with a co-purified peptide by X-ray crystallography. HhoA assembles into stable trimers, mediated by its protease domain and further into a cage-like hexamer by a novel interaction between the PDZ domains of opposing trimers. Each PDZ domain contains two loops for PDZ-PDZ formation: interaction clamp one and two (IC1, IC2). IC1 interacts with IC2 on the opposing PDZ domain and vice versa. Our structure shows a peptide bound to a conserved groove on the PDZ domain and the properties of this pocket suggest that it binds substrate proteins as well as the neo C-termini of cleaved substrates. In agreement with previous studies showing the proteolytic activity of HhoA to be activated by Ca2+ or Mg2+, binding of divalent metal ions to the central channel of the trimer by the L1 activation loop was observed.

Place, publisher, year, edition, pages
Elsevier, 2017
Keywords
sp. PCC 6803, Deg protease, HtrA protease, X-ray structure, Hexamer
National Category
Chemical Sciences
Identifiers
urn:nbn:se:umu:diva-143748 (URN)10.1016/j.jsb.2016.12.004 (DOI)000403524400001 ()
Funder
Swedish Research Council, 2011-4186Swedish Energy Agency, 2012-005889
Available from: 2018-01-08 Created: 2018-01-08 Last updated: 2018-10-15Bibliographically approved
Hall, M., Wagner, R., Lam, X. T., Funk, C. & Persson, K. (2017). The HhoA protease from Synechocystis sp. PCC 6803: novel insights into structure and activity regulation. Journal of Structural Biology, 198(3), 147-153
Open this publication in new window or tab >>The HhoA protease from Synechocystis sp. PCC 6803: novel insights into structure and activity regulation
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2017 (English)In: Journal of Structural Biology, ISSN 1047-8477, E-ISSN 1095-8657, Vol. 198, no 3, p. 147-153Article in journal (Refereed) Published
Abstract [en]

Proteases play a vital role in the removal of proteins, which become damaged due to temperature or oxidative stress. Important to this process in the cyanobacterium Synechocystis sp. PCC6803 is the family of Deg/HtrA proteases; HhoA (sll1679), HhoB (sll1427) and HtrA (slr1204). While previous studies have elucidated the structures of Deg/HtrA proteases from Escherichia coli and from the chloroplast of the higher plant Arabidopsis thaliana, no structural data have been available for any Deg/HtrA protease from cyanobacteria, the evolutionary ancestor of the chloroplast. To gain a deeper insight into the molecular mechanisms and regulation of these proteins we have solved the structure of the Synechocystis HhoA protease in complex with a co-purified peptide by X-ray crystallography. HhoA assembles into stable trimers, mediated by its protease domain and further into a cage-like hexamer by a novel interaction between the PDZ domains of opposing trimers. Each PDZ domain contains two loops for PDZ-PDZ formation: interaction clamp one and two (IC1, IC2). IC1 interacts with IC2 on the opposing PDZ domain and vice versa. Our structure shows a peptide bound to a conserved groove on the PDZ domain and the properties of this pocket suggest that it binds substrate proteins as well as the neo C-termini of cleaved substrates. In agreement with previous studies showing the proteolytic activity of HhoA to be activated by Ca2+ or Mg2+, binding of divalent metal ions to the central channel of the trimer by the L1 activation loop was observed.

Place, publisher, year, edition, pages
Elsevier, 2017
Keywords
Synechocystis sp. PCC 6803, Deg protease, HtrA protease, X-ray structure, Hexamer
National Category
Biochemistry and Molecular Biology Cell Biology Biophysics
Identifiers
urn:nbn:se:umu:diva-137441 (URN)10.1016/j.jsb.2016.12.004 (DOI)000403524400001 ()27956128 (PubMedID)
Available from: 2017-07-03 Created: 2017-07-03 Last updated: 2018-06-09Bibliographically approved
Addario, B., Sandblad, L., Persson, K. & Backman, L. (2016). Characterisation of Schizosaccharomyces pombe alpha-actinin. PeerJ, 4, Article ID e1858.
Open this publication in new window or tab >>Characterisation of Schizosaccharomyces pombe alpha-actinin
2016 (English)In: PeerJ, ISSN 2167-8359, E-ISSN 2167-8359, Vol. 4, article id e1858Article in journal (Refereed) Published
Abstract [en]

The actin cytoskeleton plays a fundamental role in eukaryotic cells. Its reorganization is regulated by a plethora of actin-modulating proteins, such as a-actinin. In higher organisms, alpha-actinin is characterized by the presence of three distinct structural domains: an N-terminal actin-binding domain and a C-terminal region with EF-hand motif separated by a central rod domain with four spectrin repeats. Sequence analysis has revealed that the central rod domain of alpha-actinin from the fission yeast Schizosaccharomyces pombe consists of only two spectrin repeats. To obtain a firmer understanding of the structure and function of this unconventional alpha-actinin, we have cloned and characterized each structural domain. Our results show that this alpha-actinin isoform is capable of forming dimers and that the rod domain is required for this. However, its actin-binding and cross-linking activity appears less efficient compared to conventional alpha-actinins. The solved crystal structure of the actin-binding domain indicates that the closed state is stabilised by hydrogen bonds and a salt bridge not present in other a-actinins, which may reduce the affinity for actin.

Keywords
Spectrin repeat, Actin-binding protein, alpha-actinin, Schizosaccharomyces pombe
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-120364 (URN)10.7717/peerj.1858 (DOI)000374159200008 ()27069798 (PubMedID)
Funder
Carl Tryggers foundation
Available from: 2016-05-17 Created: 2016-05-16 Last updated: 2018-06-07Bibliographically approved
Hasegawa, Y., Iijima, Y., Persson, K., Nagano, K., Yoshida, Y., Lamont, R., . . . Yoshimura, F. (2016). Role of Mfa5 in Expression of Mfa1 Fimbriae in Porphyromonas gingivalis. Journal of Dental Research, 95(11), 1291-1297
Open this publication in new window or tab >>Role of Mfa5 in Expression of Mfa1 Fimbriae in Porphyromonas gingivalis
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2016 (English)In: Journal of Dental Research, ISSN 0022-0345, E-ISSN 1544-0591, Vol. 95, no 11, p. 1291-1297Article in journal (Refereed) Published
Abstract [en]

Fimbriae are protein-based filamentous appendages that protrude from the bacterial cell surface and facilitate host adhesion. Two types of fimbriae, FimA and Mfa1, of the periodontal pathogen Porphyromonas gingivalis are responsible for adherence to other bacteria and to host cells in the oral cavity. Both fimbrial forms are composed of 5 proteins, but there is limited information about their polymerization mechanisms. Here, the authors evaluated the function of Mfa5, one of the Mfa1 fimbrial accessory proteins. Using mfa5 gene disruption and complementation studies, the authors revealed that Mfa5 affects the incorporation of other accessory proteins, Mfa3 and Mfa4, into fibers and the expression of fimbriae on the cell surface. Mfa5 is predicted to have a C-terminal domain (CTD) that uses the type IX secretion system (T9SS), which is limited to this organism and related Bacteroidetes species, for translocation across the outer membrane. To determine the relationship between the putative Mfa5 CTD and the T9SS, mutants were constructed with in-frame deletion of the CTD and deletion of porU, a C-terminal signal peptidase linked to T9SS-mediated secretion. The ∆CTD-expressing strain presented a similar phenotype to the mfa5 disruption mutant with reduced expression of fimbriae lacking all accessory proteins. The ∆porU mutants and the ∆CTD-expressing strain showed intracellular accumulation of Mfa5. These results indicate that Mfa5 function requires T9SS-mediated translocation across the outer membrane, which is dependent on the CTD, and subsequent incorporation into fibers. These findings suggest the presence of a novel polymerization mechanism of the P. gingivalis fimbriae.

Place, publisher, year, edition, pages
International & American Associations for Dental Research, 2016
Keywords
periodontal disease, microbiology, bacterial virulence, bacteria, biofilm, molecular biology
National Category
Chemical Sciences Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:umu:diva-124436 (URN)10.1177/0022034516655083 (DOI)000384456800012 ()27323953 (PubMedID)
Available from: 2016-08-11 Created: 2016-08-11 Last updated: 2018-06-07Bibliographically approved
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