Bacterial peptidoglycan with amidated meso-diaminopimelic acid evades NOD1 recognition: an insight into NOD1 structure-recognition

Abstract

Nucleotide-binding oligomerization domain-containing protein 1 (NOD1) is an intracellular pattern recognition receptor that recognizes bacterial peptidoglycan (PG) containing meso-diaminopimelic acid (mesoDAP) and activates the innate immune system. Interestingly, a few pathogenic and commensal bacteria modify their PG stem peptide by amidation of mesoDAP (mesoDAP_NH2). In the present study, NOD1 stimulation assays were performed using bacterial PG containing mesoDAP (PG_DAP) and mesoDAP_NH2 (PG_DAPNH2) to understand the differences in their biomolecular recognition mechanism. PG_DAP was effectively recognized, whereas PG_DAPNH2 showed reduced recognition by the NOD1 receptor. Restimulation of the NOD1 receptor, which was initially stimulated with PG_DAP using PG_DAPNH2, did not show any further NOD1 activation levels than with PG_DAP alone. But the NOD1 receptor initially stimulated with PG_DAPNH2 responded effectively to restimulation with PG_DAP The biomolecular structure-recognition relationship of the ligand-sensing leucine-rich repeat (LRR) domain of human NOD1 (NOD1-LRR) with PG_DAP and PG_DAPNH2 was studied by different computational techniques to further understand the molecular basis of our experimental observations. The d-Glu-mesoDAP motif of GMTP_DAP, which is the minimum essential motif for NOD1 activation, was found involved in specific interactions at the recognition site, but the interactions of the corresponding d-Glu-mesoDAP motif of PG_DAPNH2 occur away from the recognition site of the NOD1 receptor. Hot-spot residues identified for effective PG recognition by NOD1-LRR include W820, G821, D826 and N850, which are evolutionarily conserved across different host species. These integrated results thus successfully provided the atomic level and biochemical insights on how PGs containing mesoDAP_NH2 evade NOD1-LRR receptor recognition.

Keywords: NOD1; docking; meso-diaminopimelic acid; molecular recognition; peptidoglycan; receptor modeling.