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Infect Immun, July 1998, p. 3050-3058, Vol. 66, No. 7
School of Clinical Medical Sciences
(Rheumatology),1
Department of
Biochemistry and Genetics,2
and
School of Microbiological, Immunological and Virological
Sciences,3 The Medical School, University of
Newcastle upon Tyne, Newcastle upon Tyne, NE2 4HH, United Kingdom
Received 4 November 1997/Returned for modification 23 December
1997/Accepted 24 March 1998
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.
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Inhibition of Human Peripheral Blood Mononuclear
Cell Proliferation by Streptococcus pyogenes Cell Extract Is
Associated with Arginine Deiminase Activity
*
Corresponding author. Mailing address: Rheumatology
Laboratory, The Medical School, Framlington Place, Newcastle upon Tyne, NE2 4HH, United Kingdom. Phone: 0191 2226000, ext. 7541. Fax: 0191 2225455. E-mail: j.a.goodacre{at}ncl.ac.uk.
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