Beruflich Dokumente
Kultur Dokumente
1.
2.
3.
4.
Introduction, 80S
Oral streptococci, 80S
Carbohydrate metabolism in plaque, 81S
Fructans in dental plaque
4.1 Fructosyltransferase, 82S
4.2 Fructanase, 82S
5. Glucans in dental plaque
6.
7.
8.
9.
1. INTRODUCTION
2. ORAL STREPTOCOCCI
S TR EP T OC OC C AL SU G AR ME T AB OL I SM 81S
3. CARBOHYDRATE METABOLISM IN
PLAQUE
1997 The Society for Applied Bacteriology, Journal of Applied Microbiology Symposium Supplement 83, 80S88S
82S S .M . C O LB Y A N D R .R . B. RU S SE LL
modified
glucans
glucans
gtf C
sucrose
dexA
gtf D
+
ftf
fructose
isomaltosaccharides
fructan +
msm
glucose
fruA
dexB
fructose
glucose
modified
glucans
dei
glucans
gtf S
dexA
gtf T
sucrose
4.2 Fructanase
+
gtf U
+
isomaltosaccharides
fructose
1997 The Society for Applied Bacteriology, Journal of Applied Microbiology Symposium Supplement 83, 80S88S
S TR EP T OC OC C AL SU G AR ME T AB OL I SM 83S
Table 1 Glucosyltransferases of
Streptococcus sobrinus
5.1 Glucosyltransferase
Because of the importance of glucans in plaque, the glucosyltransferases (GTF) which make glucans have attracted
considerable interest. Streptococcus mutans produces three distinct GTF while Strep. sobrinus produces four. Each GTF is
encoded by a separate gene and each enzyme has distinctive
properties, varying in its requirement for a primer molecule
to start the polymerization reaction, the proportion of a(1
6)- and a(13)-linkages and degree of branching it introduces
into the glucan, and the total length of the glucan chain
produced. The glucans made by the individual GTF of Strep.
mutans have not been well characterized, but the GTF of
Strep. sobrinus all have individual characteristics (Table 1).
Within dental plaque, the overall properties of the glucan
present will depend on the relative activity of the different
GTF present and also on their interactions, since one GTF
may modify the product of another.
The gene sequences of 12 streptococcal GTF are now
available, as well as two from Leuconostoc mesenteroides. Multiple alignment of the deduced amino acid sequences has
revealed common features of GTF. All are of high molecular
weight (155174 kDa) and composed of distinct domains
(Fig. 2). The challenge facing those working in the field of
GTF is to identify those features of the protein structure
which determine function. We need to understand what it is
that determines why a particular GTF makes a particular
Glucosyltransferase
Glucan
Enzyme
Water
Molecular
Gene names
Primer
Structure
solubility
weight
gtf I
I* P3 Dependent
Branched,
Insoluble
High
mostly, a(13)-linkages
(insoluble)
gtf U S1 P4
Dependent
Branched,
Soluble
100 000
a(16)- and
a(13)-linkages
gtf S
S3 P2
Independent
Linear,
Soluble
5300
a(16)-linkages
gtf T
S4 P1
Independent
Branched,
Soluble
400 000
a(16)- and
a(13)-linkages
Nomenclature used by Cheetham et al. (1991)* and Hanada et al. (1993). Data on
primer dependence and glucan properties are also taken from these authors.
1997 The Society for Applied Bacteriology, Journal of Applied Microbiology Symposium Supplement 83, 80S88S
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A B
1997 The Society for Applied Bacteriology, Journal of Applied Microbiology Symposium Supplement 83, 80S88S
S TR EP T OC OC C AL SU G AR ME T AB OL I SM 85S
6. GLUCAN-BINDING PROTEINS
The glucan-binding domain of GTF may have dual functions: at least one or two of the repeat units are needed for
glucan synthesis (Ferretti et al. 1987) but the binding capacity
will also lead to glucan-mediated binding of bacterial cells
through surface-associated GTF. Free GTF is also found in
saliva and on the tooth surface where it remains active so that
glucans made on the tooth form another site of attachment
for bacteria (Schilling and Bowen 1992). Besides GTF, a
number of other glucan-binding proteins (GBP) without
known enzyme activity can be found. Streptococcus mutans
produces two GBPs, one of which consists of a series of
tandem repeat units homologous to those found in the carboxyterminal one-third of GTF (Banas et al. 1990). The
occurrence of this independent GBP is consistent with the
idea that GTF have a modular structure and this is supported
by the fact that the Dei dextranase inhibitor also contains
homologous repeat units (Sun et al. 1995). Furthermore,
homologous sequences are found in proteins from other
organisms which also have a modular structure: autolysin
of Strep. pneumoniae, fibronectin-binding protein of Strep.
pyogenes and toxin A of Clostridium difficile (Wren 1991; Talay
et al. 1994; Yother and White 1994). In all these cases the
repeat-containing module is thought to be involved in binding
to a macromolecular carbohydrate substrate, and subtle differences in protein sequence must determine the specificity
of binding.
A second GBP which induces a strong immune response
in children has been described in Strep. mutans (Smith et al.
1994) and Strep. sobrinus also produces a number of GBP (Ma
et al. 1996; Wu-Yuan and Gill 1996). Ma et al. (1996) have
1997 The Society for Applied Bacteriology, Journal of Applied Microbiology Symposium Supplement 83, 80S88S
86S S .M . C O LB Y A N D R .R . B. RU S SE LL
Streptococcus mutans and Strep. sobrinus occupy a similar ecological niche and both share the capacity to produce extracellular polymers from sucrose and to adhere to hard surfaces
by means of these polymers. Nevertheless, as Fig. 1 illustrates, the details of these processes are distinct in each species. Streptococcus mutans has three GTF and an FTF as
compared to four GTF (and no FTF) in Strep. sobrinus. While
both species produce dextranase, the controlling inhibitor Dei
is found only in Strep. sobrinus while Strep. mutans can not
only degrade dextran but also utilize the resultant oligosaccharides for intracellular metabolism. The number and
size of GBP also differs between the two species but the way in
which these are involved in adhesion is still unclear. Further
investigations of the function of all these different components should throw light on their relative contributions to
the complex process of growth and adherence in the oral
cavity. In particular, now that many of the genes have been
cloned, targeted gene inactivation allows comparison of
mutants with wild type in in vitro models of adhesion (Colby
et al. 1995) and in vivo, where the capacity to cause caries
in experimental animals can be explored (Kuramitsu 1993).
Improved knowledge of the structure of critical molecules
also sets the scene for rational design of specific inhibitors,
or identification of targets for vaccine development (Taubman
et al. 1995).
This review has presented a number of instances which
illustrate instructive parallels between research on oral streptococci and subjects in quite different areas. Examples are
the structural similarities between GTF of streptococci and
Leuconostoc mesenteroides (the commercial source of dextran),
between GTF and amylase, the finding of homologous carbohydrate binding domains in bacteria from different habitats,
and differing mechanisms of cell wall anchorage in proteins
which initially appear to have common features. Studies of
the oral streptococci thus have much to contribute to, as well
as to learn from, a wide range of microbiological systems.
8. ACKNOWLEDGEMENTS
9. REFERENCES
Banas, J.A., Russell, R.R.B. and Ferretti, J.J. (1990) Sequence
analysis of the gene for the glucan-binding protein of Streptococcus
mutans Ingbritt. Infection and Immunity 58, 667673.
Barrett, J.F., Barrett, T.A. and Curtiss, R. (1987) Purification and
partial characterization of the multicomponent dextranase complex of Streptococcus sobrinus and cloning of the dextranase gene.
Infection and Immunity 55, 792802.
Burne, R.A. and Penders, J.E.C. (1992) Characterization of the
Streptococcus mutans GS-5 fruA gene encoding exo-b-D-fructosidase. Infection and Immunity 60, 46214623.
Burne, R.A. and Penders, J.E.C. (1994) Differential localization of
the Streptococcus mutans GS-5 fructan hydrolase enzyme, FruA.
FEMS Microbiology Letters 121, 243250.
Cheetham, N.W.H., Walker, G.J., Pearce, B.J., Fiala-Beer, E. and
Taylor, C. (1991) Structures of water-soluble a-D-glucans synthesized from sucrose by glucosyltransferases isolated from Streptococcus sobrinus culture filtrates. Carbohydrate Polymers 14, 316.
Colby, S.M., Whiting, G.C., Tao, L. and Russell, R.R.B. (1995)
Insertional inactivation of the Streptococcus mutans dexA (dextranase) gene results in altered adherence and dextran catabolism.
Microbiology 141, 29292936.
Davies, G. and Henrissat, B. (1995) Structures and mechanisms of
glycosyl hydrolases. Current Opinion in Structural Biology 3, 853
859.
de Soet, J.J., Holbrook, W.P., Magnusdottir, M.O. and De Graaff,
J. (1993) Streptococcus sobrinus and Streptococcus mutans in a longitudinal study of dental caries. Microbial Ecology in Health and
Disease 6, 237243.
Dibdin, G.H. and Shellis, R.P. (1988) Physical and biochemical
studies of Streptococcus mutans sediments suggest new factors
linking the cariogenicity of plaque with its extracellular polysaccharide content. Journal of Dental Research 67, 890895.
Ebisu, S., Misaki, A., Kato, K. and Kotani, S. (1974) The structure
of water-insoluble glucans of cariogenic Streptococcus mutans, formed in the absence and presence of dextranase. Carbohydrate
Research 38, 374381.
Ellis, D.W. and Miller, C.H. (1977) Extracellular dextran hydrolase
from Streptococcus mutans strain 6715. Journal of Dental Research
56, 5769.
Ferretti, J.J., Gilpin, M.L. and Russell, R.R.B. (1987) Nucleotide
sequence of a glucosyltransferase gene from Streptococcus sobrinus
MFe28. Journal of Bacteriology 169, 42714278.
Ferretti, J.J., Russell, R.R.B. and Dao, M.L. (1989) Sequence analysis of the wall-associated protein precursor of Streptococcus mutans
antigen A. Molecular Microbiology 3, 469478.
Germaine, G.R., Harlander, S.K., Leung, W.-L.S. and Schachtele,
C.F. (1977) Streptococcus mutans dextransucrase: functioning of
primer dextran and endogenous dextranase in water-soluble and
water-insoluble glucan synthesis. Infection and Immunity 16, 637
648.
Hamelik, R.M. and McCabe, M.M. (1982) An endodextranase
inhibitor from batch cultures of Streptococcus mutans. Biochemical
and Biophysical Research Communications 106, 875880.
Hanada, N., Katayama, T., Kunimori, A., Yamashita, Y. and Takehara, T. (1993) Four different types of glucans synthesized by
glucosyltransferases from Streptococcus sobrinus. Microbios 73, 23
35.
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1997 The Society for Applied Bacteriology, Journal of Applied Microbiology Symposium Supplement 83, 80S88S