67-kDa Laminin Receptor Promotes Internalization of Cytotoxic Necrotizing Factor 1-expressing Escherichia coli K1 into Human Brain Microvascular Endothelial Cells

Kim, Kee Jun; Chung, Jin Woong; Kim, Kwang Sik

doi:10.1074/jbc.m410176200

Cited by 135Escherichia coli research studiesBacterial Infections and VaccinesBarrier Structure and Function Studies

67-kDa Laminin Receptor Promotes Internalization of Cytotoxic Necrotizing Factor 1-expressing Escherichia coli K1 into Human Brain Microvascular Endothelial Cells

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Kim, Kee Jun, Chung, Jin Woong, Kim, Kwang Sik·2005·Journal of Biological Chemistry

DOI 10.1074/jbc.m410176200

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Abstract

Escherichia coli K1 is the most common Gram-negative organism causing meningitis, and its invasion of human brain microvascular endothelial cells (HBMEC) is a prerequisite for penetration into the central nervous system. We have reported previously that cytotoxic necrotizing factor 1 (CNF1) contributes to E. coli K1 invasion of HBMEC and interacts with 37-kDa laminin receptor precursor (37LRP) of HBMEC, which is a precursor of 67-kDa laminin receptor (67LR). In the present study, we examined the role of 67LR in the CNF1-expressing E. coli K1 invasion of HBMEC. Immunofluorescence microscopy and ligand overlay assays showed that 67LR is present on the HBMEC membrane and interacts with CNF1 protein as well as the CDPGYIGSR laminin peptide. 67LR was up-regulated and clustered at the sites of E. coli K1 on HBMEC in a CNF1-dependent manner. Pretreatment of CNF1+ E. coli K1 with recombinant 37-kDa laminin receptor precursor reduced the invasion rate to the level of Deltacnf1 mutant, and the invasion rate of CNF1+ E. coli K1 was enhanced in 67LR-overexpressing HBMEC, indicating 67LR is involved in the CNF1+ E. coli K1 invasion of HBMEC. Coimmunoprecipitation analysis showed that, upon incubation with CNF1+ E. coli K1 but not with Deltacnf1 mutant, focal adhesion kinase and paxillin were recruited and associated with 67LR. When immobilized onto polystyrene beads, CNF1 was sufficient to induce internalization of coupled beads into HBMEC through interaction with 67LR. Taken together, this is the first demonstration that E. coli K1 invasion of HBMEC occurs through the ligand-receptor (CNF1-67LR) interaction, and 67LR promotes CNF1-expressing E. coli K1 internalization of HBMEC.

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Other meningeal pathogens bind ECM components and receptors to mediate bacterial-BBB interactions.
“Other meningeal pathogens, such as Streptococcus pneumoniae (SPN), E. coli K1, Neisseria meningitidis, and Haemophilus influenza type B (HiB, also bind ECM components and ECM receptors (e.g., integrins and laminin receptor) to mediate bacterial-BBB interactions (12)(13)(14)(15).”
From Bacterial induction of Snail1 contributes to blood-brain barrier disruption (2015) · cites this paper as [12]

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Cytotoxic Necrotizing Factor-1 Contributes to Escherichia coli K1 Invasion of the Central Nervous System
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Authors: Kim, K. J., Chung, J. W., & Kim, K. S.
Affiliations: Johns Hopkins University (US)
Corresponding author: Kim, Kwang Sik
Venue: Journal of Biological Chemistry
Publisher: Elsevier BV
Published: 2005-01
Volume: 280
Issue: 2
Pages: 1360-1368
ISSN: 0021-9258 (ISSN-L)
DOI: 10.1074/jbc.m410176200
OpenAlex: W1977149059
Cited by: 135
References: 29
Citations by year: 2026: 2 · 2025: 3 · 2024: 4 · 2023: 3 · 2022: 3 · 2021: 14 · 2020: 3 · 2019: 2 · 2018: 4 · 2017: 2 · 2016: 5 · 2015: 9 · 2014: 9 · 2013: 8 · 2012: 16
FWCI: 3.31
Type: journal article
Language: en
Open access: hybrid
doi.org
Licence: cc-by
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
creativecommons.org/licenses/by/4.0/ (vor, 5740 d)
Locations: Journal of Biological Chemistry (publishedVersion, cc-by)
PubMed (publishedVersion)
Funders: National Institutes of Health
National Institute of Allergy and Infectious Diseases (AI-47225, R01AI047225)
National Institute of Neurological Disorders and Stroke (NS-26310)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Endocrinology › Escherichia coli research studies
Life Sciences › Immunology and Microbiology › Microbiology › Bacterial Infections and Vaccines
Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
Keywords: 67-kDa laminin receptor, cytotoxic necrotizing factor 1, Escherichia coli K1, meningitis, brain microvascular endothelial cells, bacterial invasion, receptor internalization, focal adhesion kinase, paxillin, ligand-receptor interactions, laminin peptides, blood-brain barrier
MeSH terms: Bacterial Toxins, Binding Sites, Brain, Cells, Cultured, Cerebrovascular Circulation, Cytoskeletal Proteins, Escherichia coli, Humans, Lysosomes, Microcirculation, Microspheres, Molecular Weight, Phosphoproteins, Protein-Tyrosine Kinases, Up-Regulation, Receptors, Laminin, Escherichia coli Proteins, Endothelial Cells, Immunoprecipitation, Focal Adhesion Protein-Tyrosine Kinases, Focal Adhesion Kinase 1, Paxillin

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67-kDa Laminin Receptor Promotes Internalization of Cytotoxic Necrotizing Factor 1-expressing Escherichia coli K1 into Human Brain Microvascular Endothelial Cells

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