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

doi:10.1016/j.celrep.2023.112515

Cell-surface anchoring of Listeria adhesion protein on L. monocytogenes is fastened by internalin B for pathogenesis

Cell Rep. 2023 May 30;42(5):112515. doi: 10.1016/j.celrep.2023.112515. Epub 2023 May 11.

Authors

Dongqi Liu¹, Xingjian Bai², Harrison D B Helmick³, Manalee Samaddar¹, Mary Anne Roshni Amalaradjou⁴, Xilin Li², Shivendra Tenguria¹, Nicholas L F Gallina¹, Luping Xu³, Rishi Drolia⁵, Uma K Aryal⁶, Gustavo Marçal Schmidt Garcia Moreira⁷, Michael Hust⁷, Mohamed N Seleem⁸, Jozef L Kokini³, Raluca Ostafe⁹, Abigail Cox¹⁰, Arun K Bhunia¹¹

Affiliations

¹ 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.
² Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN, USA.
³ Department of Food Science, Purdue University, West Lafayette, IN, USA.
⁴ Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN, USA; Department of Animal Sciences, University of Connecticut, Storrs, CT, USA.
⁵ 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.
⁶ Bindley Bioscience, Purdue University, West Lafayette, IN, USA; Department of Comparative Pathobiology, Purdue University, West Lafayette, IN, USA.
⁷ Technische Universität Braunschweig University of Technology, Institute for Biochemistry, Biotechnology, and Bioinformatics, Spielmannstr. 7, 38106 Braunschweig, Germany.
⁸ 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.
⁹ Purdue Institute of Inflammation, Immunology, and Infectious Disease, West Lafayette, IN, USA.
¹⁰ Department of Comparative Pathobiology, Purdue University, West Lafayette, IN, USA.
¹¹ 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. Electronic address: bhunia@purdue.edu.

PMID: 37171960
DOI: 10.1016/j.celrep.2023.112515

Abstract

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 (K_D ∼ 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.

Keywords: AdhE; CP: Microbiology; InlB; LAP; Listeria adhesion protein; Listeria monocytogenes; acetaldehyde alcohol dehydrogenase; bacterial translocation; neuroinvasion; pathogenesis; protein-protein interaction; secretion; surface anchoring.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alcohol Dehydrogenase / metabolism
Animals
Bacterial Proteins / metabolism
Cell Membrane / metabolism
Listeria monocytogenes* / metabolism
Listeria* / metabolism
Membrane Proteins / metabolism
Mice

Substances

Bacterial Proteins
Membrane Proteins
Alcohol Dehydrogenase