A unique network of attack, defence and competence on the outer membrane of the periodontitis pathogen <i>Tannerella forsythia</i>

Książek, Mirosław; Goulas, Theodoros; Mizgalska, Danuta; Rodríguez-Banqueri, Arturo; Eckhard, Ulrich; Veillard, Florian; Waligorska, Irena; Benedyk-Machaczka, Małgorzata; Sochaj-Gregorczyk, Alicja; Madej, Mariusz; Thøgersen, Ida B.; Enghild, Jan J.; Cuppari, Anna; Arolas, Joan L.; Diego, Iñaki de; López-Pelegrín, Mar; Garcia-Ferrer, I.; Guevara, Tibisay; Dive, Vincent; Zani, Marie-Louise; Moreau, Thierry; Potempa, Jan; Gomis‐Rüth, F. Xavier

doi:10.1039/d2sc04166a

Cited by 3 publications

(4 citation statements)

References 109 publications

(213 reference statements)

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“…2D ). We also tested mutants deficient for the secreted KLIKK proteases karilysin and mirolysin, and the two cell‐surface protease inhibitors miropin and potempin A [ 19 , 23 ]. Interestingly, the mirolysin‐deficient mutant showed a significant decrease in binding affinity for both aptamers, with the MFI falling to 11.35% for TF1 and 7.9% for TF2 compared to the wild‐type strain (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The T. forsythia mutant deficient for the S‐layer was obtained by homologous recombination as previously described [ 19 ]. Briefly, vector pTfsAB_LONG_cat_pUC57 (containing a chloramphenicol resistance cassette flanked by an 845‐bp sequence upstream of the TfsA start codon and a 1274‐bp sequence downstream of the TfsB stop codon) was synthesized by GenScript Biotech, Piscataway, NJ, USA.…”

Section: Methodsmentioning

confidence: 99%

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Screening and characterization of aptamers recognizing the periodontal pathogen Tannerella forsythia

Mizgalska,

Malicki,

Golda

et al. 2024

FEBS Open Bio

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Periodontal disease is one of the most common forms of inflammation. It is currently diagnosed by observing symptoms such as gingival bleeding and attachment loss. However, the detection of biomarkers that precede such symptoms would allow earlier diagnosis and prevention. Aptamers are short oligonucleotides or peptides that fold into three‐dimensional conformations conferring the ability to bind molecular targets with high affinity and specificity. Here we report the selection of aptamers that bind specifically to the bacterium Tannerella forsythia, a pathogen frequently associated with periodontal disease. Two aptamers with the highest affinity were examined in more detail, revealing that their binding is probably dependent on mirolysin, a surface‐associated protease secreted by the T. forsythia type‐9 secretion system. The aptamers showed minimal cross‐reactivity to other periodontopathogens and are therefore promising leads for the development of new tools to study the composition of the periodontitis‐associated dysbiotic bacteriome as well as inexpensive new diagnostic assays.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Screening and characterization of aptamers recognizing the periodontal pathogen Tannerella forsythia

Mizgalska,

Malicki,

Golda

et al. 2024

FEBS Open Bio

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“…Overall, the moiety is cohered by an internal hydrophobic core made up by 23 residues ( figure 3 f ). Finally, the C-terminal segment of strand β7 encompasses the sequence K 787 –L–I–K–K 791 , which is found at the C-terminus of five unique serine- and metal-dependent peptidases secreted by the T9SS of T. forsythia in addition to mirolase, collectively named ‘KLIKK-peptidases’ [ 65 , 74 ]. This conservation is in line with the above sequence requirements of motif C-t. in P. gingivalis, which also includes a conserved lysine in the middle of C-terminal strand β7 (see §2.2).…”

Section: Resultsmentioning

confidence: 99%

“…Here, we sought to identify molecular determinants for T9SS secretion and PorU-cleavage in P. gingivalis by comparing the four aforementioned structures with two ad hoc high-confidence homology models and verified them with a cohort of mutant strains that were subjected to cellular and functional assays. Furthermore, we determined the crystal structure of the CTD of the T. forsythia T9SS cargo mirolase [ 65 ] (dubbed ‘LKK’; from L 706 to K–K 791 , see UniProt entry (UP) A0A0A7KVG3 for residue numbering), the first outside P. gingivalis , and assessed if the above structural determinants were conserved across species. Finally, we checked the CTD compatibility of T. forsythia and P. gingivalis .…”

Section: Introductionmentioning

confidence: 99%

Structural and functional insights into the C-terminal signal domain of the Bacteroidetes type-IX secretion system

Mizgalska,

Rodríguez-Banqueri,

Veillard

et al. 2024

Open Biol.

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Gram-negative bacteria from the Bacteroidota phylum possess a type-IX secretion system (T9SS) for protein secretion, which requires cargoes to have a C-terminal domain (CTD). Structurally analysed CTDs are from Porphyromonas gingivalis proteins RgpB, HBP35, PorU and PorZ, which share a compact immunoglobulin-like antiparallel 3+4 β-sandwich (β1–β7). This architecture is essential as a P. gingivalis strain with a single-point mutant of RgpB disrupting the interaction of the CTD with its preceding domain prevented secretion of the protein. Next, we identified the C-terminus (‘motif C-t.’) and the loop connecting strands β3 and β4 (‘motif Lβ3β4’) as conserved. We generated two strains with insertion and replacement mutants of PorU, as well as three strains with ablation and point mutants of RgpB, which revealed both motifs to be relevant for T9SS function. Furthermore, we determined the crystal structure of the CTD of mirolase, a cargo of the Tannerella forsythia T9SS , which shares the same general topology as in Porphyromonas CTDs. However, motif Lβ3β4 was not conserved. Consistently, P. gingivalis could not properly secrete a chimaeric protein with the CTD of peptidylarginine deiminase replaced with this foreign CTD. Thus, the incompatibility of the CTDs between these species prevents potential interference between their T9SSs.

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Integrating Large-Scale Protein Structure Prediction into Human Genetics Research

Correa Marrero,

Jänes,

Baptista

et al. 2024

Annual Review of Genomics and Human Genetics

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The last five years have seen impressive progress in deep learning models applied to protein research. Most notably, sequence-based structure predictions have seen transformative gains in the form of AlphaFold2 and related approaches. Millions of missense protein variants in the human population lack annotations, and these computational methods are a valuable means to prioritize variants for further analysis. Here, we review the recent progress in deep learning models applied to the prediction of protein structure and protein variants, with particular emphasis on their implications for human genetics and health. Improved prediction of protein structures facilitates annotations of the impact of variants on protein stability, protein–protein interaction interfaces, and small-molecule binding pockets. Moreover, it contributes to the study of host–pathogen interactions and the characterization of protein function. As genome sequencing in large cohorts becomes increasingly prevalent, we believe that better integration of state-of-the-art protein informatics technologies into human genetics research is of paramount importance.

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A unique network of attack, defence and competence on the outer membrane of the periodontitis pathogen Tannerella forsythia

Abstract: Periodontopathogenic Tannerella forsythia uniquely secretes six peptidases of disparate catalytic classes and families that operate as virulence factors during infection of the gums, the KLIKK-peptidases. Their coding genes are immediately...

Cited by 3 publications

References 109 publications

Screening and characterization of aptamers recognizing the periodontal pathogen Tannerella forsythia

Screening and characterization of aptamers recognizing the periodontal pathogen Tannerella forsythia

Structural and functional insights into the C-terminal signal domain of the Bacteroidetes type-IX secretion system

Integrating Large-Scale Protein Structure Prediction into Human Genetics Research

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