Bacterial peptide recognition and immune activation facilitated by human peptide transporter PEPT2.
Microbial detection requires the recognition of pathogen-associated molecular patterns (PAMPs) by pattern recognition receptors (PRRs) that are distributed on the cell surface and within the cytosol. The nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) family functions as an intracellular PRR that triggers the innate immune response. The mechanism by which PAMPs enter the cytosol to interact with NLRs, particularly muropeptides derived from the bacterial proteoglycan cell wall, is poorly understood. PEPT2 is a proton-dependent transporter that mediates the active translocation of di- and tripeptides across epithelial tissues, including the lung. Using computational tools, we initially established that bacterial dipeptides, particularly gamma-D-glutamyl-meso-diaminopimelic acid (gamma-iE-DAP), are suitable substrates for PEPT2. We then determined in primary cultures of human upper airway epithelia and transiently transfected CHO-PEPT2 cell lines that gamma-iE-DAP uptake was mediated by PEPT2 with an affinity constant of approximately 193 microM, whereas muramyl dipeptide was not transported. Exposure to gamma-iE-DAP at the apical surface of differentiated, polarized cultures resulted in activation of the innate immune response in an NOD1- and RIP2-dependent manner, resulting in release of IL-6 and IL-8. Based on these findings we report that PEPT2 plays a vital role in microbial recognition by NLR proteins, particularly with regard to airborne pathogens, thereby participating in host defense in the lung.
Swaan
Peter W
PW
Department of Pharmaceutical Sciences, University of Maryland, Baltimore, MD, USA.
Bensman
Timothy
T
Bahadduri
Praveen M
PM
Hall
Mark W
MW
Sarkar
Anasuya
A
Bao
Shengying
S
Khantwal
Chandra M
CM
Ekins
Sean
S
Knoell
Daren L
DL
D000818:Animals; D001426:Bacterial Proteins / Q000494:pharmacology*; D002478:Cells, Cultured; D003198:Computer Simulation; D006224:Cricetinae; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007113:Immunity, Innate / Q000276:immunology; D008168:Lung / Q000187:drug effects / Q000378:metabolism; D008958:Models, Molecular; D015394:Molecular Structure; D053474:Nod1 Signaling Adaptor Protein / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000494:pharmacology*; D011485:Protein Binding; D021381:Protein Transport; D053475:Receptor-Interacting Protein Serine-Threonine Kinase 2 / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D027981:Symporters / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*
D001426:Bacterial Proteins; D053474:Nod1 Signaling Adaptor Protein; D010455:Peptides; D027981:Symporters; C093694:hydrogen-coupled oligopeptide transporter PepT2; D053475:Receptor-Interacting Protein Serine-Threonine Kinase 2