Cell-surface anchoring of Listeria adhesion protein on L. monocytogenes is fastened by internalin B for pathogenesis
Dongqi Liu,
Xingjian Bai,
Harrison D.B. Helmick,
Manalee Samaddar,
Mary Anne Roshni Amalaradjou,
Xilin Li,
Shivendra Tenguria,
Nicholas L.F. Gallina,
Luping Xu,
Rishi Drolia,
Uma K. Aryal,
Gustavo Marçal Schmidt Garcia Moreira,
Michael Hust,
Mohamed N. Seleem,
Jozef L. Kokini,
Raluca Ostafe,
Abigail Cox,
Arun K. Bhunia
Affiliations
Dongqi Liu
Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN, USA; Purdue Institute of Inflammation, Immunology, and Infectious Disease, West Lafayette, IN, USA
Xingjian Bai
Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN, USA
Harrison D.B. Helmick
Department of Food Science, Purdue University, West Lafayette, IN, USA
Manalee Samaddar
Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN, USA; Purdue Institute of Inflammation, Immunology, and Infectious Disease, West Lafayette, IN, USA
Mary Anne Roshni Amalaradjou
Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN, USA; Department of Animal Sciences, University of Connecticut, Storrs, CT, USA
Xilin Li
Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN, USA
Shivendra Tenguria
Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN, USA; Purdue Institute of Inflammation, Immunology, and Infectious Disease, West Lafayette, IN, USA
Nicholas L.F. Gallina
Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN, USA; Purdue Institute of Inflammation, Immunology, and Infectious Disease, West Lafayette, IN, USA
Luping Xu
Department of Food Science, Purdue University, West Lafayette, IN, USA
Rishi Drolia
Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN, USA; Purdue Institute of Inflammation, Immunology, and Infectious Disease, West Lafayette, IN, USA; Department of Biological Science, Eastern Kentucky University, Richmond, KY, USA
Uma K. Aryal
Bindley Bioscience, Purdue University, West Lafayette, IN, USA; Department of Comparative Pathobiology, Purdue University, West Lafayette, IN, USA
Gustavo Marçal Schmidt Garcia Moreira
Technische Universität Braunschweig University of Technology, Institute for Biochemistry, Biotechnology, and Bioinformatics, Spielmannstr. 7, 38106 Braunschweig, Germany
Michael Hust
Technische Universität Braunschweig University of Technology, Institute for Biochemistry, Biotechnology, and Bioinformatics, Spielmannstr. 7, 38106 Braunschweig, Germany
Mohamed N. Seleem
Department of Comparative Pathobiology, Purdue University, West Lafayette, IN, USA; Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA
Jozef L. Kokini
Department of Food Science, Purdue University, West Lafayette, IN, USA
Raluca Ostafe
Purdue Institute of Inflammation, Immunology, and Infectious Disease, West Lafayette, IN, USA
Abigail Cox
Department of Comparative Pathobiology, Purdue University, West Lafayette, IN, USA
Arun K. Bhunia
Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN, USA; Purdue Institute of Inflammation, Immunology, and Infectious Disease, West Lafayette, IN, USA; Department of Comparative Pathobiology, Purdue University, West Lafayette, IN, USA; Corresponding author
Summary: Listeria adhesion protein (LAP) is a secreted acetaldehyde alcohol dehydrogenase (AdhE) that anchors to an unknown molecule on the Listeria monocytogenes (Lm) surface, which is critical for its intestinal epithelium crossing. In the present work, immunoprecipitation and mass spectrometry identify internalin B (InlB) as the primary ligand of LAP (KD ∼ 42 nM). InlB-deleted and naturally InlB-deficient Lm strains show reduced LAP-InlB interaction and LAP-mediated pathology in the murine intestine and brain invasion. InlB-overexpressing non-pathogenic Listeria innocua also displays LAP-InlB interplay. In silico predictions reveal that a pocket region in the C-terminal domain of tetrameric LAP is the binding site for InlB. LAP variants containing mutations in negatively charged (E523S, E621S) amino acids in the C terminus confirm altered binding conformations and weaker affinity for InlB. InlB transforms the housekeeping enzyme, AdhE (LAP), into a moonlighting pathogenic factor by fastening on the cell surface.
