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Infection and Immunity, August 2000, p. 4604-4610, Vol. 68, No. 8
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Differential Protein Expression in Phenotypic
Variants of Streptococcus pneumoniae
Karin
Overweg,1
Chris D.
Pericone,2
Gerridina
G. C.
Verhoef,1
Jeffrey N.
Weiser,2
Hugo D.
Meiring,3
Ad P. J. M.
De
Jong,3
Ronald
De
Groot,1 and
Peter W. M.
Hermans1,*
Department of Pediatrics, Sophia Children's
Hospital, Erasmus University, Rotterdam,1 and
Laboratory of Organic Analytical Chemistry, National Institute
of Public Health and the Environment,
Bilthoven,3 The Netherlands, and
Departments of Pediatrics and Microbiology, University of
Pennsylvania School of Medicine, Philadelphia,
Pennsylvania2
Received 8 December 1999/Returned for modification 20 March
2000/Accepted 26 April 2000
Streptococcus pneumoniae undergoes spontaneous phase
variation resulting in opaque and transparent colony forms. Differences in colony opacity correlate with differences in virulence: the transparent variants are more capable of colonizing the nasopharynx, whereas the opaque variants show increased virulence during systemic infections. To gain insight into the pathogenesis of pneumococcal disease at the molecular level, protein expression patterns of the
phenotypic variants of two pneumococcal strains were compared by
high-resolution two-dimensional protein electrophoresis. In comparison
with transparent variants, the opaque variants reduced the expression
of two proteins and overexpressed one protein. The proteins were
identified by mass spectrometric analysis. The protein overexpressed in
the opaque phenotype revealed significant homology to elongation factor
Ts of Helicobacter pylori. One of the two proteins that
were underexpressed in the opaque variants revealed significant
homology to the proteinase maturation protein PrtM of
Lactocobacillus paracasei, a member of the family of
peptidyl-prolyl cis/trans isomerases. A consensus
lipoprotein signal sequence suggests that the putative proteinase
maturation protein A, designated PpmA, is located at the surface of the
pneumococcus and may play a role in the maturation of surface or
secreted proteins. The second underexpressed protein was identified as
pyruvate oxidase, SpxB. The lower SpxB expression in opaque variants
most probably explains the reduced production of hydrogen peroxide, a
reaction product of SpxB, in this variant. Since a
spxB-defective pneumococcal mutant has decreased ability to
colonize the nasopharynx (B. Spellerberg, D. R. Cundell, J. Sandros, B. J. Pearce, I. Idanpaan-Heikkila, C. Rosenow, and
H. R. Masure, 1996. Mol. Microbiol. 19:803-813, 1996), our data
suggest that SpxB plays an important role in enhancing the ability of
transparent variants to efficiently colonize the nasopharynx.
*
Corresponding author. Mailing address: Laboratory of
Pediatrics, Room Ee 1500, Erasmus University Rotterdam, P.O. Box 1738, 3000 DR Rotterdam, The Netherlands. Phone: 31-10-4088224. Fax: 31-10-4089486. E-mail: hermans{at}kgk.fgg.eur.nl.
Infection and Immunity, August 2000, p. 4604-4610, Vol. 68, No. 8
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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