BUSCADOR RECOLECTA

Raw microscopy data for Fig3B, 3E and 3S1 (.lif format) in a single zip file., Resources available on the publisher's site: https://doi.org/10.25452/figshare.plus.18309440.v1, Ministry of Science, Innovation and Universities BFU2017-83562-P, Peer reviewed

Raw microscopy data for Fig5E (.lif files) in a single zip file., Resources available on the publisher's site: https://doi.org/10.25452/figshare.plus.18348926.v1, Marie Skłodowska-Curie grant agreement No. 754510 Ministry of Science, Innovation and Universities PGC2018-095616-B-I00 Ministry of Science, Innovation and Universities PGC2018-099562-B-I00, Peer reviewed

Resources available on the publisher's site: https://doi.org/10.5281/zenodo.5554758, Streptococcal pyrogenic exotoxin B (SpeB) is a cysteine protease expressed during group A streptococcal infection that represents a major virulence factor. Although subject to several studies, its role during infection is still under debate, and its proteolytic properties remain insufficiently characterized. Here, we revisited this protease through a set of complementary approaches relying on state of-the-art HPLC-MS methods. After conceiving an efficient protocol to recombinantly express SpeB, the zymogen of the protease and its activation were characterized. Employing proteome-derived peptide libraries, a strong preference for hydrophobic and aromatic residues at P2 alongside negatively charged amino acids at P3′ to P6′ was revealed. To identify relevant in vivo substrates, native proteins were obtained from monocytic secretome and plasma to assess their cleavage under physiological conditions. Besides corroborating our findings concerning specificity, more than 200 cleaved proteins were identified, including proteins of the extracellular matrix, proteins of the immune system, and proteins involved in inflammation. Finally, the cleavage of IgG subclasses was studied in detail. This study precisely depicts the proteolytic properties of SpeB and provides a library of potential host substrates, including their exact cleavage positions, as a valuable source for further research to unravel the role of SpeB during streptococcal infection., Peer reviewed

Additional file 1: Table ST1. Treatments comparison by outcome in male patients. Table ST2. Treatments comparison by outcome in female patients. Table ST3. WHO classification of disease outcome. Table ST4. Panels and antibodies used for immunophenotyping. Figure SF1. Patients distribution by outcome, age, and comorbidities. Figure SF2. Distribution of male patients with comorbidities according to age and testosterone levels. Figure SF3. Longitudinal analysis of serum levels of IL-6, C-reactive protein (CRP), ferritin and lactate dehydrogenase (LDH) in male patients. Figure SF4. Bioavailable testosterone serum levels and correlation between age and sex-hormone binding globulin (SHBG). Figure SF5. Flow cytometry analysis of circulating immune subpopulations in three illustrative cases with moderate, severe survivor and severe deceased outcomes., Ministerio de Ciencia, Innovación y Universidades Consejo Superior de Investigaciones Científicas Agència de Gestió d’Ajuts Universitaris i de Recerca Conselleria d'Educació, Investigació, Cultura i Esport Instituto de Salud Carlos III, Peer reviewed

Additional file 2 Figure S1. Percentage of G-Actin in filaments. Percentage of G-Actin in filaments of different size for the three characteristic regimes., Ministerio de Ciencia, Innovación y Universidades Ministerio de Ciencia, Innovación y Universidades, Peer reviewed

Additional file 3 Figure S2. Dependence of speed of treadmiling with F-actin length. Plot of the dependence of the instantaneous speed on the filament size. Short filaments move faster than average. Long filaments move at the same speed in average., Ministerio de Ciencia, Innovación y Universidades, Peer reviewed

Additional file 6: Movie S3. Time-lapse movie of the system showing oscillations. Time-lapse movie of the system showing periodic assembly and disassembly of the actomyosin cortex (G-actin labeled in green, ACs labeled in red, Myosin labeled in red)., Ministerio de Ciencia, Innovación y Universidades, Peer reviewed

Additional file 7 Figure S4. Oscillations for different levels of Myosin. Oscillations in the number of G-actin in the cortex for conditions of (A) low and (B) high concentration of Myosin in the system. The corresponding Fourier transform for each oscillation is also shown., Ministerio de Ciencia, Innovación y Universidades, Peer reviewed

Additional file 8: Movie S4. Oscillations in D. Melanogaster basal follicle cells. Control conditions. Cells are stained with Lifeact-GFP., Ministerio de Ciencia, Innovación y Universidades, Peer reviewed

Additional file 12 Figure S7. Scheme of the effect of Myosin over F-actin. (1), Myosin movement in parallel f-Actin. (2) F-actin sliding. (3) Tension building in the filaments. (4) Release from cortex after threshold tension is reached. After release, tension (illustrated in red) is redistributed to other F-actin in the network., Ministerio de Ciencia, Innovación y Universidades, Peer reviewed