Inhibition of Bacterial Peptidoglycan Cytopathy by Retina Pigment Epithelial PGRP2 Amidase

Adv Exp Med Biol. 2023:1415:521-526. doi: 10.1007/978-3-031-27681-1_76.

Abstract

Peptidoglycan (PGN) recognition protein 2 (PGRP2; N-acetylmuramyl-L-alanine amidase (NAMAA)) activity in corneal epithelial cells is thought to inhibit corneal inflammation by reducing the PGN-induced cytokines. PGRP2 has not been reported in human retinal pigment epithelial (RPE) cells. RPE cell lysate NAMAA activity was measured densitometrically via cleavage of FITC-tagged muramyl dipeptide (FITCMDP). RPE lysate degradation of the cytopathic activity of nucleotide-binding oligomerization domain (NOD) receptor agonists was assessed by caspase-3 activation and DNA ladder detection and quantitation. PGRP2/NAMAA protein was detected in RPE cells by immunofluorescent antibody assay. RPE lysate NAMAA cleaved FITCMDP in a dose- and time-dependent manner. RPE lysate selectively inhibited PGN cytopathic activity of NOD1 agonists containing D-γ-glutamyl-meso-diaminopimelic acid and NOD2 containing L-alanyl-D-isoglutamine. The results suggest RPE PGRP2 amidase selectively degrades PGN that stimulate NOD-mediated cytopathic activity. The failure of RPE NAMAA to degrade pro-inflammatory PGN may play a role in bacterial retinopathies.

Keywords: Bacteria; Cytopathy; Meso-diaminopimelic acid; Muramyl dipeptide; N-acetylmuramyl-L-alanine amidase; NOD agonist; Retinopathy.

MeSH terms

  • Acetylmuramyl-Alanyl-Isoglutamine / metabolism
  • Acetylmuramyl-Alanyl-Isoglutamine / pharmacology
  • Amidohydrolases / metabolism
  • Cytokines* / metabolism
  • Fluorescein-5-isothiocyanate
  • Humans
  • Nod2 Signaling Adaptor Protein / metabolism
  • Peptidoglycan* / chemistry
  • Peptidoglycan* / metabolism
  • Retina / metabolism

Substances

  • Peptidoglycan
  • Fluorescein-5-isothiocyanate
  • Cytokines
  • Acetylmuramyl-Alanyl-Isoglutamine
  • Amidohydrolases
  • Nod2 Signaling Adaptor Protein