Localization and mutagenesis of the sorting signal binding site on sortase A from Staphylococcus aureus.
Surface proteins in Gram-positive bacteria are anchored to the cell wall by the action of sortase enzymes. The Staphylococcus aureus sortase A (SrtA) protein anchors proteins by recognizing a cell wall sorting signal containing the amino acid sequence LPXTG. To understand how SrtA binds this sequence, we carried out NMR studies of new peptidyl-cyanoalkene and peptidyl-sulfhydryl inhibitors that contain the sorting signal sequence LPAT. These studies combined with amino acid mutagenesis identified a catalytically important and conserved binding surface formed by residues A118, T180, and I182. Compatible with its recently proposed role as a general base, R197 is also shown to be required for catalysis.
Liew
Chu Kong
CK
Department of Chemistry and Biochemistry, University of California Los Angeles, 405 Hilgard Avenue, Los Angeles, CA 90095-1570, USA.
Smith
Brenton T
BT
Pilpa
Rosemarie
R
Suree
Nuttee
N
Ilangovan
Udayar
U
Connolly
Kevin M
KM
Jung
Michael E
ME
Clubb
Robert T
RT
D000595:Amino Acid Sequence; D019881:Aminoacyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins; D001665:Binding Sites; D002473:Cell Wall / Q000201:enzymology; D003546:Cysteine Endopeptidases; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D011487:Protein Conformation; D015398:Signal Transduction; D013211:Staphylococcus aureus / Q000201:enzymology* / Q000235:genetics
D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases