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Selected Publication:

Jarosch, M; Egelseer, EM; Huber, C; Moll, D; Mattanovich, D; Sleytr, UB; 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. 2001; 147(Pt 5):1353-1363

Abstract:
The mature surface layer (S-layer) protein SbsC of Bacillus stearothermophilus ATCC 12980 comprises amino acids 31-1099 and self-assembles into an oblique lattice type which functions as an adhesion site for a cell-associated high-molecular-mass exoamylase. To elucidate the structure-function relationship of distinct segments of SbsC, three N- and seven C-terminal truncations were produced in a heterologous expression system, isolated, purified and their properties compared with those of the recombinant mature S-layer protein rSbsC(31-1099). With the various truncated forms it could be demonstrated that the N-terminal part (aa 31-257) is responsible for anchoring the S-layer subunits via a distinct type of secondary cell wall polymer to the rigid cell wall layer, but this positively charged segment is not required for the self-assembly of SbsC, nor for generating the oblique lattice structure. If present, the N-terminal part leads to the formation of in vitro double-layer self-assembly products. Affinity studies further showed that the N-terminal part includes an exoamylase-binding site. Interestingly, the N-terminal part carries two sequences of 6 and 7 aa (AKAALD and KAAYEAA) that were also identified on the amylase-binding protein AbpA of Streptococcus gordonii, In contrast to the self-assembling N-terminal truncation rSbsC(258-1099), two further N-terminal truncations (rSbsC(343-1099), rSbsC(447-1099)) and three C-terminal truncations (rSbsC(31-713), rSbsC(31-844), rSbsC(31-860)) had lost the ability to self-assemble and stayed in the water-soluble state, Studies with the self-assembling C-terminal truncations rSbsC(31-880), rSbsC(31-900), and rSbsC(31-920) revealed that the C-terminal 219 aa can be deleted without interfering with the self-assembly process, while the C-terminal 179 aa are not required for the formation of the oblique lattice structure.
Authors BOKU Wien:
Egelseer Eva Maria
Mattanovich Diethard
Sleytr Uwe B.
BOKU Gendermonitor:

Find related publications in this database (using NML MeSH Indexing)
Bacillus stearothermophilus - chemistry
Bacterial Proteins - chemistry
Cell Wall - metabolism
Chromatography, Affinity - metabolism
Electrophoresis, Polyacrylamide Gel - metabolism
Escherichia coli - genetics
Membrane Glycoproteins - chemistry
Microscopy, Electron - chemistry
Molecular Sequence Data - chemistry
Peptidoglycan - chemistry
Polymerase Chain Reaction - chemistry
Protein Binding - chemistry
Protein Structure, Tertiary - chemistry
Recombinant Proteins - genetics
Sequence Deletion - genetics
Structure-Activity Relationship - genetics

Find related publications in this database (Keywords)
heterologous expression
self assembly
peptidoglycan
secondary cell wall polymer
exoamylase


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