title: Biochemical and biological activity of arginine deiminase from Streptococcus pyogenes M22.
abstract: Streptococcus pyogenes (group A Streptococcus; GAS) is an important gram-positive extracellular bacterial pathogen responsible for a number of suppurative infections. This micro-organism has developed complex virulence mechanisms to avoid the host's defenses. We have previously reported that SDSC from GAS type M22 causes endothelial-cell dysfunction, and inhibits cell adhesion, migration, metabolism, and proliferation in a dose-dependent manner, without affecting cell viability. This work aimed to isolate and characterize a component from GAS type M22 supernatant that suppresses the proliferation of endothelial cells (EA.hy926). In the process of isolating a protein possessing antiproliferative activity we identified arginine deiminase (AD). Further study showed that this enzyme is most active at pH 6.8. Calculating Km and Vmax gave the values of 0.67 mmol·L(-1) and 42 s(-1), respectively. A distinctive feature of AD purified from GAS type M22 is that its optimum activity and the maximal rate of the catalytic process is close to neutral pH by comparison with enzymes from other micro-organisms. AD from GAS type M22 suppressed the proliferative activity of endothelial cells in a dose-dependent mode. At the same time, in the presence of AD, the proportion of cells in G0/G1 phase increased. When l-Arg was added at increasing concentrations to the culture medium containing AD (3 μg·mL(-1)), the enzyme's capacity to inhibit cell proliferation became partially depressed. The proportion of cells in phases S/G2 increased concomitantly, although the cells did not fully recover their proliferation activity. This suggests that AD from GAS type M22 has potential for the suppression of excessive cell proliferation.
authors.affiliation: a Institute of Experimental Medicine, 12 Pavlov Street, St. Petersburg, 197376, Russia.
authors.forename: Eleonora A
authors.initials: EA
authors.lastname: Starikova
authors.affiliation: a Institute of Experimental Medicine, 12 Pavlov Street, St. Petersburg, 197376, Russia.|b Saint-Petersburg State University, 7-9 Universitetskaya nab., St. Petersburg, 199034, Russia.
authors.forename: Alexey V
authors.initials: AV
authors.lastname: Sokolov
authors.affiliation: a Institute of Experimental Medicine, 12 Pavlov Street, St. Petersburg, 197376, Russia.
authors.forename: Anna Yu
authors.initials: AY
authors.lastname: Vlasenko
authors.affiliation: a Institute of Experimental Medicine, 12 Pavlov Street, St. Petersburg, 197376, Russia.
authors.forename: Larisa A
authors.initials: LA
authors.lastname: Burova
authors.affiliation: a Institute of Experimental Medicine, 12 Pavlov Street, St. Petersburg, 197376, Russia.
authors.forename: Irina S
authors.initials: IS
authors.lastname: Freidlin
authors.affiliation: a Institute of Experimental Medicine, 12 Pavlov Street, St. Petersburg, 197376, Russia.|b Saint-Petersburg State University, 7-9 Universitetskaya nab., St. Petersburg, 199034, Russia.
authors.forename: Vadim B
authors.initials: VB
authors.lastname: Vasilyev
mesh_terms: D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000378:metabolism / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006867:Hydrolases / Q000378:metabolism*; D013297:Streptococcus pyogenes / Q000201:enzymology*; D013329:Structure-Activity Relationship
chemical_list: D017077:Culture Media, Conditioned; D006867:Hydrolases; C019723:arginine deiminase
keywords: Streptococcus pyogenes M22; arginine deiminase; arginine déiminase; endothelial cell proliferation; prolifération des cellules endothéliales