Inhibition of human peripheral blood mononuclear cell proliferation by Streptococcus pyogenes cell extract is associated with arginine deiminase activity.
Streptococcus pyogenes (group A Streptococcus) cell extracts (CE) have a remarkably powerful and dose-dependent inhibitory effect on antigen, superantigen, or mitogen-stimulated human peripheral blood mononuclear cell (PBMC) proliferation in vitro. Purification of the inhibitory component present in S. pyogenes type M5 (Manfredo strain) CE by anion-exchange chromatography followed by gel filtration chromatography showed that the inhibitor had an approximate native molecular mass of 100 kDa. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified inhibitory fractions followed by silver staining gave a single band with an approximate molecular mass of 47 kDa, indicating that the inhibitor is composed of two identical subunits. NH2-terminal sequencing of the protein revealed that it was identical to the previously characterized streptococcal acid glycoprotein (SAGP); this protein possesses between 31.5 and 39.0% amino acid identity with arginine deiminase (AD) from Mycoplasma hominis, Mycoplasma arginini, Pseudomonas putida, and Pseudomonas aeruginosa. AD enzyme activity was present in unfractionated CE prepared from a range of streptococcal strains, and partially purified inhibitory fractions of Manfredo CE also had high levels of activity. The inhibitory effect of Manfredo CE was overcome by the addition of L-arginine to proliferation assays in which human PBMC were stimulated with phytohemagglutinin. We conclude that SAGP, or its homolog, possesses AD activity and that the potent inhibition of proliferation of human T cells by streptococcal CE is due to activity of this enzyme.
Degnan
B A
BA
School of Clinical Medical Sciences (Rheumatology), Immunological and Virological Sciences, The Medical School, University of Newcastle upon Tyne, Newcastle upon Tyne, NE2 4HH, United Kingdom.
Palmer
J M
JM
Robson
T
T
Jones
C E
CE
Fischer
M
M
Glanville
M
M
Mellor
G D
GD
Diamond
A G
AG
Kehoe
M A
MA
Goodacre
J A
JA
D000595:Amino Acid Sequence; D001120:Arginine / Q000494:pharmacology; D015551:Autoimmunity; D006801:Humans; D006867:Hydrolases / Q000502:physiology*; D007166:Immunosuppressive Agents; D008213:Lymphocyte Activation; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013297:Streptococcus pyogenes / Q000737:chemistry / Q000276:immunology / Q000502:physiology*; D013601:T-Lymphocytes / Q000276:immunology*
D007166:Immunosuppressive Agents; D001120:Arginine; D006867:Hydrolases; C019723:arginine deiminase