In vitro reconstitution demonstrates the cell wall ligase activity of LCP proteins.
Sacculus is a peptidoglycan (PG) matrix that protects bacteria from osmotic lysis. In Gram-positive organisms, the sacculus is densely functionalized with glycopolymers important for survival, but the way in which assembly occurs is not known. In Staphylococcus aureus, three LCP (LytR-CpsA-Psr) family members have been implicated in attaching the major glycopolymer wall teichoic acid (WTA) to PG, but ligase activity has not been demonstrated for these or any other LCP proteins. Using WTA and PG substrates produced chemoenzymatically, we show that all three proteins can transfer WTA precursors to nascent PGs, establishing that LCP proteins are PG-glycopolymer ligases. Although all S. aureus LCP proteins have the capacity to attach WTA to PG, we show that their cellular functions are not redundant. Strains lacking lcpA have phenotypes similar to those of WTA-null strains, indicating that this is the most important WTA ligase. This work provides a foundation for studying how LCP enzymes participate in cell wall assembly.
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.|Department of Microbiology and Immunology, Harvard Medical School, Boston, Massachusetts, USA.
Kaitlin
K
Schaefer
Department of Microbiology and Immunology, Harvard Medical School, Boston, Massachusetts, USA.
Leigh M
LM
Matano
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.|Department of Microbiology and Immunology, Harvard Medical School, Boston, Massachusetts, USA.
Yuan
Y
Qiao
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
Daniel
D
Kahne
Department of Microbiology and Immunology, Harvard Medical School, Boston, Massachusetts, USA.
Suzanne
S
Walker
D001426:Bacterial Proteins / Q000378:metabolism*; D002473:Cell Wall / Q000201:enzymology* / Q000378:metabolism*; D066298:In Vitro Techniques; D008025:Ligases / Q000378:metabolism*; D015394:Molecular Structure; D010457:Peptidoglycan / Q000737:chemistry / Q000378:metabolism; D013211:Staphylococcus aureus / Q000166:cytology* / Q000201:enzymology* / Q000378:metabolism
D001426:Bacterial Proteins; D010457:Peptidoglycan; D008025:Ligases