TY - JOUR
T1 - Structure of the LdcB LD-carboxypeptidase reveals the molecular basis of peptidoglycan recognition
AU - Hoyland, Christopher N.
AU - Aldridge, Christine
AU - Cleverley, Robert M.
AU - Duchêne, Marie Clémence
AU - Minasov, George
AU - Onopriyenko, Olena
AU - Sidiq, Karzan
AU - Stogios, Peter J.
AU - Anderson, Wayne F.
AU - Daniel, Richard A.
AU - Savchenko, Alexei
AU - Vollmer, Waldemar
AU - Lewis, Richard J.
N1 - Funding Information:
We thank the staff at the Diamond Synchrotron Light Source for access to, and help with, its beamlines for X-ray diffraction data collection. We thank Arnaud Baslé, Joseph Newman, and Vincent Rao for useful discussions; Atikah Mohd Sukor for excellent technical assistance; Z. Wawrzak, S. Shatsman, and S.N. Peterson for assistance with data collection and refinement; and T. Skarina for assistance with purification and crystallization. We also thank the authors of Meziane-Cherif et al. (2014) for sharing results and atomic coordinates prior to publication. Finally, we thank Joe Gray of the Newcastle University Pinnacle-Proteomics and Biological Mass Spectrometry facility for mass spectrometry analysis of muropeptide fractions. This project has been funded in whole or in part with federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, U.S. Department of Health and Human Services (contracts HHSN272200700058C and HHSN2722012000026C), and by the UK Biotechnology and Biological Sciences Research Council in an award to R.A.D., W.V., and R.J.L. (BB/G015902/1).
PY - 2014/7/8
Y1 - 2014/7/8
N2 - Peptidoglycan surrounds the bacterial cytoplasmic membrane to protect the cell against osmolysis. The biosynthesis of peptidoglycan, made of glycan strands crosslinked by short peptides, is the target of antibiotics like β-lactams and glycopeptides. Nascent peptidoglycan contains pentapeptides that are trimmed by carboxypeptidases to tetra- and tripeptides. The well-characterized DD-carboxypeptidases hydrolyze the terminal D-alanine from the stem pentapeptide to produce a tetrapeptide. However, few LD-carboxypeptidases that produce tripeptides have been identified, and nothing is known about substrate specificity in these enzymes. We report biochemical properties and crystal structures of the LD-carboxypeptidases LdcB from Streptococcus pneumoniae, Bacillus anthracis, and Bacillus subtilis. The enzymes are active against bacterial cell wall tetrapeptides and adopt a zinc-carboxypeptidase fold characteristic of the LAS superfamily. We have also solved the structure of S. pneumoniae LdcB with a product mimic, elucidating the residues essential for peptidoglycan recognition and the conformational changes that occur on ligand binding.
AB - Peptidoglycan surrounds the bacterial cytoplasmic membrane to protect the cell against osmolysis. The biosynthesis of peptidoglycan, made of glycan strands crosslinked by short peptides, is the target of antibiotics like β-lactams and glycopeptides. Nascent peptidoglycan contains pentapeptides that are trimmed by carboxypeptidases to tetra- and tripeptides. The well-characterized DD-carboxypeptidases hydrolyze the terminal D-alanine from the stem pentapeptide to produce a tetrapeptide. However, few LD-carboxypeptidases that produce tripeptides have been identified, and nothing is known about substrate specificity in these enzymes. We report biochemical properties and crystal structures of the LD-carboxypeptidases LdcB from Streptococcus pneumoniae, Bacillus anthracis, and Bacillus subtilis. The enzymes are active against bacterial cell wall tetrapeptides and adopt a zinc-carboxypeptidase fold characteristic of the LAS superfamily. We have also solved the structure of S. pneumoniae LdcB with a product mimic, elucidating the residues essential for peptidoglycan recognition and the conformational changes that occur on ligand binding.
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U2 - 10.1016/j.str.2014.04.015
DO - 10.1016/j.str.2014.04.015
M3 - Article
C2 - 24909784
AN - SCOPUS:84904103328
VL - 22
SP - 949
EP - 960
JO - Structure with Folding & design
JF - Structure with Folding & design
SN - 0969-2126
IS - 7
ER -