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Previous Article | Next Article 
Journal of Bacteriology, December 1999, p. 7643-7646, Vol. 181, No. 24
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Structural and Functional Analyses of the Secondary Cell Wall
Polymer of Bacillus sphaericus CCM 2177 That Serves as
an S-Layer-Specific Anchor
Nicola
Ilk,1
Paul
Kosma,2
Michael
Puchberger,2
Eva M.
Egelseer,1
Harald F.
Mayer,1
Uwe B.
Sleytr,1 and
Margit
Sára1,*
Zentrum für Ultrastrukturforschung und
Ludwig Boltzmann-Institut für Molekulare
Nanotechnologie1 and Institut
für Chemie,2 University of Agricultural
Sciences, 1180 Vienna, Austria
Received 15 July 1999/Accepted 4 October 1999
 |
ABSTRACT |
Sacculi of Bacillus sphaericus CCM 2177 contain a
secondary cell wall polymer which was completely extracted with
48% hydrofluoric acid. Nuclear magnetic resonance analysis showed that
the polymer is composed of repeating units, as follows:
3)-[4,6-O-(1-carboxyethylidene)]~0.5-
-D-ManpNAc-(14)--D-GlcpNAc-(1. The N-terminal part of the S-layer protein carrying S-layer homologous motifs recognizes this polymer as a binding site.
| |
TEXT |
Crystalline bacterial cell surface
layers (S-layers) represent the outermost cell envelope component of
many bacteria and archaea (for reviews, see references 4, 23,
27, and 28). S-layers are composed of
identical proteinaceous subunits, and they assemble into either
oblique, square, or hexagonal lattice types. To answer the
question of how S-layer proteins of gram-positive bacteria are anchored
to the rigid cell wall layer, the whole cell envelope complexes of
Bacillus stearothermophilus wild-type strains and an
oxygen-induced variant strain were analyzed, and secondary cell wall
polymers (SCWP) were found to function as binding sites for this class
of secreted proteins (9, 21). Based on structure,
teichoic acids, teichuronic acids, lipoteichoic acids, and lipoglycans
are distinguished among SCWP (for reviews, see references 3,
10, 19, and 22). Most of the biological functions ascribed to SCWP, such as binding of cations, protecting the
cell against toxic metals, keeping the peptidoglycan sacculus in an
expanded state by charge repulsion, binding of protons to create an
acidic cell wall during bacterial growth, and providing a biophysical
barrier to prevent diffusion of substances, have been viewed in the
context of their acidic nature (for a review, see reference
3).
In contrast to those of B. stearothermophilus wild-type
strains (9, 11), the S-layer proteins of most gram-positive
bacteria carry three typical S-layer homologous (SLH) motifs
(16) at the N-terminal part, each of them consisting of
approximately 50 to 60 amino acids. In addition to being present in
S-layer proteins, SLH motifs were also identified at the C-terminal end of cell-associated exoenzymes or other exoproteins (14, 17). In several studies, SLH motifs were found to anchor the different types
of cell-associated exoproteins to the rigid cell wall layer (6, 7,
14, 15, 17, 18, 20). However, only for a few organisms was it
confirmed that an SCWP is involved in the binding process (6, 7,
18, 21). In the present study, the structure of the SCWP of
Bacillus sphaericus CCM 2177 was characterized by nuclear
magnetic resonance (NMR) analysis. Moreover, evidence was provided that
the SCWP, not the peptidoglycan, recognizes the N-terminal part of the
S-layer protein carrying at least one SLH motif.
Characterization of the S-layer of B. sphaericus CCM
2177.
Freeze-etching of whole cells from B. sphaericus
CCM 2177 grown in continuous culture in nutrient broth at 30°C at
a dilution rate of 0.16 h
1 revealed that the cell
surface was completely covered with a square S-layer lattice showing a
rather smooth outer surface (data not shown). The S-layer protein could
completely be extracted from cell wall fragments prepared as described
previously (9, 21, 25) with 5 M guanidine hydrochloride
(GHCl). During removal of GHCl by dialysis against 10 mM
CaCl2 at 20°C, the S-layer subunits assembled into flat
sheets with a maximum width of 2 µm. Negative staining and
freeze-drying revealed that the self-assembly products represented
double layers in which the individual S-layers were oriented to each
other, with corrugated inner surfaces (Fig.
1). Upon dialysis of the GHCl-extracted
S-layer protein against distilled water or 10 mM EDTA at 20°C, mainly
amorphous aggregates were observed in negatively stained preparations
(data not shown). The positive effect of calcium ions on the in vitro
self-assembly was also seen by the degree of assembly (percentage of
total S-layer protein assembled [21]). When dialysis
was performed against distilled water or 10 mM EDTA, the degree of
assembly was <5% after 4 h and increased to 25% 18 h after
the dialysis procedure was started. For comparison, degrees of assembly
of 55 and 80%, respectively, were achieved at the above-mentioned
points of time when dialysis was performed against 10 mM
CaCl2.
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FIG. 1.
Electron micrograph of a freeze-dried S-layer
self-assembly product of B. sphaericus CCM 2177. The outer
S-layer surface is rather smooth, whereas the inner S-layer surface is
much more corrugated. Bar, 200 nm.
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|
Chemical analyses of native and HF-extracted
peptidoglycan-containing sacculi.
Native peptidoglycan-containing
sacculi and those extracted with 48% HF were prepared as described
previously (21). The results from amino acid and amino sugar
analyses of hydrolyzed samples of native and HF-extracted sacculi are
summarized in Table 1. For calculating
the molar ratios between the individual components, glutamic acid (Glu)
was set to a value of 1. With the exception of the excess
glucosamine (GlcNH2) and the occurrence of
substantial amounts of mannosamine (ManNH2), the
molar ratios of all other components were typical of the
A4
-chemotype (26). Extracting the native
peptidoglycan-containing sacculi with 48% HF for 48 h at 4°C
(21) led to the complete removal of the excess of
GlcNH2 and of ManNH2, whereas the molar ratios
for all other peptidoglycan constituents remained unchanged (Table 1).
In comparison to native peptidoglycan-containing sacculi, those
extracted with HF did not show any changes in size and morphology but
appeared less electron dense in negatively stained preparations (Fig.
2a and d).
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FIG. 2.
Electron micrographs of negatively stained and
ultrathin-sectioned preparations of native and HF-extracted
peptidoglycan-containing sacculi of B. sphaericus
CCM 2177. (a and b) Negatively stained preparations of native
peptidoglycan-containing sacculi before and after recrystallization of
the S-layer protein, respectively; (c) ultrathin-sectioned preparation
of native peptidoglycan-containing sacculi after recrystallization of
the S-layer protein; (d and e) negatively stained preparations of
HF-extracted peptidoglycan-containing sacculi (48% HF, 48 h at
4°C) before and after the addition of the GHCl-extracted S-layer
protein and dialysis, respectively; (f) ultrathin-sectioned preparation
of HF-extracted peptidoglycan-containing sacculi after the addition of
the GHCl-extracted S-layer protein and dialysis. Bars, 250 nm.
|
|
Chemical and NMR analysis of the HF-extracted purified SCWP.
The HF-extracted SCWP, which represented about 50% of the native
peptidoglycan-containing sacculi by weight, was purified by gel
permeation chromatography according to a method described previously (24). Hydrolysis of the SCWP with 4 N HCl (6 h at 110°C) led to a GlcNH2-to-ManNH2
molar ratio of 1 to 1, whereas a molar ratio of 2.1 to 1 was
obtained by using 6 N HCl (6 h at 110°C). For NMR analysis, 4.6 mg of the purified SCWP was dissolved in D2O (0.6 ml,
99.95%). Spectra were recorded at 300 and 330 K at 300.13 MHz
for 1H and at 75.47 MHz for 13C with a
Bruker AVANCE 300 spectrometer equipped with a 5-mm
QNP probehead with z gradients. 1H spectra were referenced
internally to sodium 3-trimethylsilyl-1-propane sulfonate (
= 0); 13C spectra were referenced externally to 1,4-dioxane
( = 67.40). COSY, TOCSY, HMQC, HMBC, and NOESY spectra were
recorded with standard XWINNMR software (Bruker). 1H and
13C NMR spectra recorded at 300 K revealed the presence of
pyruvate groups as indicated by a signal at 1.50 ppm in the proton
domain and by signals at 24.79, ~100.0, and 174.9 ppm in the HMBC
spectrum. The signal intensity of the pyruvate methyl groups
corresponded to ~20 to 25% of the integral of the neighboring
N-acetyl groups at 2.02 to 2.04. Pyruvate substituents were cleaved
off, and a well-resolved spectrum was recorded at 330 K (Table
2). The 1H NMR spectrum
displayed two major signals for anomeric protons at 4.82 ppm
(J ~ 1.0 Hz) and 4.59 ppm (J ~ 7.7 Hz) as well as two minor signals at 4.88 and 5.21 ppm. Since the
13C NMR spectrum contained only two major signals for
anomeric carbons (100.55 and 98.89 ppm), the SCWP had to be composed of
disaccharide repeating units occurring in the -anomeric
configuration (JC1 = 164.2 and 165.1 Hz,
respectively). The two minor anomeric 1H and
13C signals were assigned to reducing -configured
N-acetyl mannosamine (ManNAc; 5.21 and 91.8 ppm) and terminal
2-acetamido-2-deoxy--D-hexopyranosyl units (4.88 and
100.2 ppm). Comparison of the proton signal intensities indicated an
average of 8 to 9 disaccharide residues in the polysaccharide sample.
This finding was substantiated by matrix-assisted laser desorption
ionization-time of flight data which revealed major signals at
2,073.4, 2,887.2, 3,294.2, 3,700.9 and 4,107.5 mass units,
corresponding to 6 to 10 N-acetyl hexosamine disaccharide units [(HexNAc)2] units (M + Na + H2O).
Homonuclear and heteronuclear correlation spectra allowed the
straightforward assignment of two units of
2-acetamido-2-deoxy-mannopyranosyl and
2-acetamido-2-deoxy-glucopyranosyl residues. NOESY spectra revealed an
interresidue Noe from H-1 of the N-acetyl glucosamine (GlcNAc) residues to H-3 of the ManNAc units, whereas irradiation of
H-1 of the ManNAc moieties yielded signal enhancement of H-3 and -4 (having similar chemical shifts) of the GlcNAc units. Since substitution at C-3 would lead to an upfield shift of C-2, the observed
shift value for C-2 (56.14 ppm) was only compatible with a
4-O-substitution of the GlcNAc unit. The observed value for the optical
rotation []D20 
16° (c 0.4, H2O) of the SCWP indicated the presence of two units each
of -D-configured residues of ManNAc and GlcNAc
(12). Thus, the structure of the pyruvic acid-free SCWP may
be proposed to be as follows:
3)--D-ManpNAc-(14)--D-GlcpNAc-(1.
Recrystallization and affinity studies with whole S-layer
protein and proteolytic cleavage fragments as well as
native and HF-extracted peptidoglycan-containing
sacculi.
The GHCl-extracted S-layer protein was mixed
with native or HF-extracted sacculi under conditions described
previously (21), and the suspensions were dialyzed
against distilled water, 10 mM CaCl2, or 10 mM EDTA at
4°C for 18 h. Ultrathin sectioning revealed that complete
outer and inner S-layers were formed on native peptidoglycan-containing
sacculi when dialysis was performed against distilled water or 10 mM
CaCl2 (Fig. 2c). Negative staining showed that the
formation of an extensive square lattice structure was strongly
dependent on the presence of calcium ions during dialysis (Fig. 2a and
b). If dialysis was against distilled water, only small, randomly
oriented patches with square lattice symmetry consisting of up to 10 morphological units were formed (data not shown). When dialysis was
performed against 10 mM EDTA, neither the square S-layer lattice nor
the outer or inner S-layer could be observed (data not shown). Thus,
the results from electron microscopic investigations demonstrated that
calcium ions are required for the correct binding of the S-layer
protein to the rigid cell wall layer as well as for the formation of
the square lattice structure. Independent of the dialysis conditions,
the S-layer protein did not bind to HF-extracted (48% HF, 48 h,
4°C) sacculi (Fig. 2d to f), which, according to chemical analysis, represented pure peptidoglycan (Table 1).
S-layer self-assembly products were dissolved in 2 M GHCl, and the
S-layer protein was cleaved with endoproteinase Glu-C under conditions
described previously (9, 21, 24). After removing GHCl by
dialysis against 50 mM Tris-HCl buffer (pH 7.2), native peptidoglycan-containing sacculi were added. When the mixture was
centrifuged, uncleaved S-layer protein and three major protein bands
with estimated relative molecular masses of 57,000, 38,000, and 32,000 Da were enriched in the pellet (Fig. 3),
whereas all other cleavage fragments remained unbound. The protein
bands which showed affinity to native sacculi had N termini (AQVND)
identical to that of the whole S-layer protein
(AQVNDYNKISGYAKEAVQSLVDQGVIOGDTNGNFN) (SLH motif underlined) showing an estimated relative molecular mass of
127,000. Distinct protein bands of those remaining in the clear
supernatant showing apparent relative molecular masses of 53,000, 28,000, and 25,000 Da were also subjected to N-terminal sequencing.
Their N-terminal regions were TAPNG, DVKNT, and TAPNG, none of
which could be identified within the N-terminal sequence of the whole
S-layer protein. None of the proteolytic cleavage fragments could
bind to HF-extracted sacculi (Fig. 3) representing pure peptidoglycan
(Table 1).
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FIG. 3.
Lanes: a, SDS-PAGE pattern obtained by proteolytic
degradation of the whole S-layer protein of B. sphaericus
CCM 2177 with endoproteinase Gluc-C; b and e, proteolytic cleavage
fragments that remained in the clear supernatant after incubation with
native and HF-extracted peptidoglycan-containing sacculi, respectively;
c and f, proteolytic cleavage fragments which could be removed from the
bound fraction by washing native and HF-extracted
peptidoglycan-containing sacculi, respectively, with buffer; d and g,
proteolytic cleavage fragments which remained attached to native and
HF-extracted peptidoglycan-containing sacculi, respectively. Molecular
masses (in thousands) are shown at the left. Protein bands subjected to
N-terminal sequencing are indicated by arrowheads.
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|
To conclude, the N-terminal part of the S-layer protein from B. sphaericus CCM 2177, carrying at least one SLH motif,
recognizes a net negatively charged SCWP composed of GlcNAc,
ManNAc, and pyruvic acid as the binding site. According to
the chemical composition, this SCWP should be attributed to the
teichuronic acids (3). The basic structure of the SCWP of
B. sphaericus CCM 2177 is similar to those of
polysaccharides from Bacillus polymyxa AHU 1385 (13), type e capsular polysaccharide of Haemophilus
influenzae (29), and glycans copurified with the
S-layer glycoproteins of Thermoanaerobacterium thermosaccharolyticum E207-71 and D120-70 (1, 2). The
N-terminal 35 amino acids of the S-layer protein from B. sphaericus CCM 2177 showed 97.1 and 88.6% identity to the
corresponding N-terminal parts of the S-layer proteins from
B. sphaericus P-1 and 2362 (5, 8). The
S-layer proteins from B. sphaericus P-1 and 2362 showed
80% identity for their N-terminal regions, while the sequence identity
beyond the N-terminal 200 amino acids was less than 20%. Accordingly,
the internal cleavage fragments of the S-layer protein of B. sphaericus CCM 2177 could not be mapped on the sequences
of the S-layer proteins from B. sphaericus P-1 and 2362.
| |
ACKNOWLEDGMENTS |
This work was supported by the Austrian Science Foundation (project
P 12938), by the Ministry of Science and Transports, and by
Hochschuljubiläumsstiftung der Stadt Wien (project H121/98).
We thank Christoph Hotzy and Aida Medovic for excellent technical
assistance, Sonja Zayni for amino acid and sugar analyses, and Fritz
Altmann for providing the matrix-assisted laser desorption ionization data.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Zentrum
für Ultrastrukturforschung, Universität für
Bodenkultur, Gregor-Mendelstr. 33, 1180 Vienna, Austria. Phone:
0043-1-47654/2208. Fax: 0043-1-478 91 12. E-mail:
sara{at}edv1.boku.ac.at.
| |
REFERENCES |
| 1.
|
Altman, E.,
J.-R. Brisson,
P. Messner, and U. B. Sleytr.
1990.
Chemical characterization of the regularly arranged surface layer glycoprotein of Clostridium thermosaccharolyticum D120-70.
Eur. J. Biochem.
188:73-82[Medline].
|
| 2.
|
Altman, E.,
C. Schäffer,
J.-R. Brisson, and P. Messner.
1996.
Isolation and characterization of an amino sugar-rich glycopeptide from the surface layer glycoprotein of Clostridium thermosaccharolyticum D120-70.
Carbohydr. Res.
295:245-253[Medline].
|
| 3.
|
Archibald, A. R.,
I. C. Hancock, and C. R. Harwood.
1993.
Cell wall structure, synthesis, and turnover, p. 381-410.
In
A. Sonenshein, J. A. Hoch, and R. Losick (ed.), Bacillus subtilis and other gram-positive bacteria. Academic Press, New York, N.Y
|
| 4.
|
Beveridge, T. J.
1994.
Bacterial S-layers.
Curr. Opin. Struct. Biol.
4:204-212.
|
| 5.
|
Bowditch, R. D.,
P. Baumann, and A. A. Yousten.
1989.
Cloning and sequencing of the gene encoding a 125-kilodalton surface-layer protein from Bacillus sphaericus 2362 and of a related cryptic gene.
J. Bacteriol.
171:4178-4188[Abstract/Free Full Text].
|
| 6.
|
Brechtel, E., and H. Bahl.
1999.
In Thermoanaerobacterium thermosulfurigenes EM1 S-layer homology domains do not attach to peptidoglycan.
J. Bacteriol.
181:5017-5023[Abstract/Free Full Text].
|
| 7.
|
Chauvaux, S.,
M. Matuschek, and P. Béguin.
1999.
Distinct affinity of binding sites for S-layer homologous domains in Clostridium thermocellum and Bacillus anthracis cell envelopes.
J. Bacteriol.
181:2455-2458[Abstract/Free Full Text].
|
| 8.
|
Deblaere, R.
July 1995.
Expression of surface layer proteins. Patent WO 9519371-A
|
| 9.
|
Egelseer, E. M.,
K. Leitner,
M. Jarosch,
C. Hotzy,
S. Zayni,
U. B. Sleytr, and M. Sára.
1998.
The S-layer proteins of two Bacillus stearothermophilus wild-type strains are bound via their N-terminal region to a secondary cell wall polymer of identical chemical composition.
J. Bacteriol.
180:1488-1495[Abstract/Free Full Text].
|
| 10.
|
Fischer, W.
1994.
Lipoteichioic acids and lipoglycans, p. 199-216.
In
J.-M. Ghuysen, and R. Hakenbeck (ed.), Bacterial cell wall. Elsevier, Amsterdam, The Netherlands
|
| 11.
| Jarosch, M., E. M. Egelseer, D. Mattanovich,
U. B. Sleytr, and M. Sára. The S-layer gene
sbsC of Bacillus stearothermophilus ATCC 12980:
molecular characterization and heterologous expression in
Escherichia coli. Microbiology, in press.
|
| 12.
|
Klyne, W.
1950.
The configuration of the anomeric carbon atoms in some cardiac glycosides.
Biochem. J.
47:xli-xlii.
|
| 13.
|
Kojima, N.,
S. Kaya,
Y. Araki, and E. Ito.
1988.
Pyruvic-acid-containing polysaccharide in the cell wall of Bacillus polymyxa AHU 1385.
Eur. J. Biochem.
174:255-260[Medline].
|
| 14.
|
Leibovitz, E.,
M. Lemaire,
I. Miras,
S. Salamitou,
P. Béguin,
H. Ohayon,
P. Gounon,
M. Matuschek,
K. Sahm, and H. Bahl.
1997.
Occurrence and function of a common domain in S-layer and other exocellular proteins.
FEMS Microbiol. Rev.
20:127-133.
|
| 15.
|
Lemaire, M.,
I. Miras,
P. Gounon, and P. Béguin.
1998.
Identification of a region responsible for binding to a cell wall within the S-layer protein of Clostridium thermocellum.
Microbiology
144:211-217[Abstract].
|
| 16.
|
Lupas, A.
1996.
A circular permutation event in the evolution of the SLH domain?
Mol. Microbiol.
20:897-898[Medline].
|
| 17.
|
Matuschek, M.,
G. Burchhardt,
K. Sahm, and H. Bahl.
1994.
Pullulanase of Thermoanaerobacterium thermosulfurigenes EM1 (Clostridium thermosulfurogenes): molecular analysis of the gene, composite structure of the enzyme, and a common model for its attachment to the cell surface.
J. Bacteriol.
176:3295-3302[Abstract/Free Full Text].
|
| 18.
|
Mesnage, S.,
E. Tosi-Couture, and A. Fouet.
1999.
Production and cell surface anchoring of functional fusion proteins between the SLH motifs of the Bacillus anthracis S-layer proteins and the Bacillus subtilis levansucrase.
Mol. Microbiol.
31:927-936[Medline].
|
| 19.
|
Navarre, W. W., and O. Schneewind.
1999.
Surface proteins of gram-positive bacteria and mechanisms of their targeting to the cell wall envelope.
Microbiol. Mol. Biol. Rev.
63:174-229[Abstract/Free Full Text].
|
| 20.
|
Olabarria, G.,
J. L. Carrascosa,
M. A. de Pedro, and J. Berenguer.
1996.
A conserved motif in S-layer proteins is involved in peptidoglycan binding in Thermus thermophilus.
J. Bacteriol.
178:4765-4772[Abstract/Free Full Text].
|
| 21.
|
Ries, W. C.,
Hotzy,
I. Schocher,
U. B. Sleytr, and M. Sára.
1997.
Evidence that a secondary cell wall polymer recognizes the N-terminal part of the S-layer protein from Bacillus stearothermophilus PV72/p2.
J. Bacteriol.
179:3892-3898[Abstract/Free Full Text].
|
| 22.
|
Salton, M. R. J.
1994.
The bacterial cell envelope a historical perspective, p. 1-22.
In
J.-M. Ghuysen, and R. Hakenbeck (ed.), Bacterial cell wall. Elsevier, Amsterdam, The Netherlands
|
| 23.
|
Sára, M., and U. B. Sleytr.
1996.
Crystalline bacterial cell surface layers (S-layers): from cell structure to biomimetics.
Prog. Biophys. Mol. Biol.
65:83-111[Medline].
|
| 24.
|
Sára, M.,
C. Dekitsch,
H. F. Mayer,
E. M. Egelseer, and U. B. Sleytr.
1998.
Influence of the secondary cell wall polymer on the reassembly, recrystallization, and stability properties of the S-layer protein from Bacillus stearothermophilus PV72/p2.
J. Bacteriol.
180:4146-4153[Abstract/Free Full Text].
|
| 25.
|
Sára, M.,
B. Kuen,
H. F. Mayer,
F. Mandl,
K. C. Schuster, and U. B. Sleytr.
1996.
Dynamics in oxygen-induced changes in S-layer protein synthesis from Bacillus stearothermophilus PV72 and its S-layer-deficient variant T5 in continuous culture and studies of the cell wall composition.
J. Bacteriol.
178:2108-2117[Abstract/Free Full Text].
|
| 26.
|
Schleifer, K. H., and O. Kandler.
1972.
Peptidoglycan types of bacterial cell walls and their taxonomic implications.
Bacteriol. Rev.
36:407-477[Free Full Text].
|
| 27.
|
Sleytr, U. B., and T. J. Beveridge.
1999.
Bacterial S-layers.
Trends Microbiol.
7:253-260[Medline].
|
| 28.
|
Sleytr, U. B.,
P. Messner,
D. Pum, and M. Sára.
1999.
Crystalline bacterial cell surface layers (S-layers): from supramolecular cell structure to biomimetics and nanotechnology.
Angew. Chem. Int. Ed.
38:1034-1054.
|
| 29.
|
Tsui, F.-P.,
R. Schneerson,
R. A. Boykins,
A. B. Karpas, and W. Egan.
1981.
Structural and immunological studies of the Haemophilus influenzae type d capsular polysaccharide.
Carbohydr. Res.
97:293-306[Medline].
|
Journal of Bacteriology, December 1999, p. 7643-7646, Vol. 181, No. 24
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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Runzler, D., Huber, C., Moll, D., Kohler, G., Sara, M.
(2004). Biophysical Characterization of the Entire Bacterial Surface Layer Protein SbsB and Its Two Distinct Functional Domains. J. Biol. Chem.
279: 5207-5215
[Abstract]
[Full Text]
-
Calabi, E., Fairweather, N.
(2002). Patterns of Sequence Conservation in the S-Layer Proteins and Related Sequences in Clostridium difficile. J. Bacteriol.
184: 3886-3897
[Abstract]
[Full Text]
-
Ilk, N., Vollenkle, C., Egelseer, E. M., Breitwieser, A., Sleytr, U. B., Sara, M.
(2002). Molecular Characterization of the S-Layer Gene, sbpA, of Bacillus sphaericus CCM 2177 and Production of a Functional S-Layer Fusion Protein with the Ability To Recrystallize in a Defined Orientation while Presenting the Fused Allergen. Appl. Environ. Microbiol.
68: 3251-3260
[Abstract]
[Full Text]
-
Karjalainen, T., Waligora-Dupriet, A.-J., Cerquetti, M., Spigaglia, P., Maggioni, A., Mauri, P., Mastrantonio, P.
(2001). Molecular and Genomic Analysis of Genes Encoding Surface-Anchored Proteins from Clostridium difficile. Infect. Immun.
69: 3442-3446
[Abstract]
[Full Text]
-
Jarosch, M., Egelseer, E. M., Huber, C., Moll, D., Mattanovich, D., Sleytr, U. B., Sára, M.
(2001). Analysis of the structure-function relationship of the S-layer protein SbsC of Bacillus stearothermophilus ATCC 12980 by producing truncated forms. Microbiology
147: 1353-1363
[Abstract]
[Full Text]
-
Martínez, B., Sillanpää, J., Smit, E., Korhonen, T. K., Pouwels, P. H.
(2000). Expression of cbsA Encoding the Collagen-Binding S-Protein of Lactobacillus crispatus JCM5810 in Lactobacillus casei ATCC 393T. J. Bacteriol.
182: 6857-6861
[Abstract]
[Full Text]
-
Sára, M., Sleytr, U. B.
(2000). S-Layer Proteins. J. Bacteriol.
182: 859-868
[Full Text]
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