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Functional analysis of the sortase YhcS in Bacillus subtilis
Author(s) -
Fasehee Hamidreza,
Westers Helga,
Bolhuis Albert,
Antelmann Haike,
Hecker Michael,
Quax Wim J.,
Mirlohi Agha F.,
van Dijl Jan Maareten,
Ahmadian Gholamreza
Publication year - 2011
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201100174
Subject(s) - sortase , bacillus subtilis , sortase a , cell wall , biology , biochemistry , mutant , microbiology and biotechnology , bacteria , bacterial protein , genetics , gene
Sortases of Gram‐positive bacteria catalyze the covalent C‐terminal anchoring of proteins to the cell wall. Bacillus subtilis , a well‐known host organism for protein production, contains two putative sortases named YhcS and YwpE. The present studies were aimed at investigating the possible sortase function of these proteins in B. subtilis . Proteomics analyses revealed that sortase‐mutant cells released elevated levels of the putative sortase substrate YfkN into the culture medium upon phosphate starvation. The results indicate that YfkN required sortase activity of YhcS for retention in the cell wall. To analyze sortase function in more detail, we focused attention on the potential sortase substrate YhcR, which is co‐expressed with the sortase YhcS. Our results showed that the sortase recognition and cell‐wall‐anchoring motif of YhcR is functional when fused to the Bacillus pumilus chitinase ChiS, a readily detectable reporter protein that is normally secreted. The ChiS fusion protein is displayed at the cell wall surface when YhcS is co‐expressed. In the absence of YhcS, or when no cell‐wall‐anchoring motif is fused to ChiS, the ChiS accumulates predominately in the culture medium. Taken together, these novel findings show that B. subtilis has a functional sortase for anchoring proteins to the cell wall.