Pseudomonas aeruginosa uses multiple receptors for adherence to laminin during infection of the respiratory tract and skin wounds

Abstract

Pseudomonas aeruginosa efficiently adheres to human tissues, including the lungs and skin, causing infections that are difficult to treat. Laminin is a main component of the extracellular matrix, and in this study we defined bacterial laminin receptors on P. aeruginosa. Persistent clinical P. aeruginosa isolates from patients with cystic fibrosis, wounds or catheter-related urinary tract infections bound more laminin compared to blood isolates. Laminin receptors in the outer membrane were revealed by 2D-immunblotting, and the specificities of interactions were confirmed with ELISA and biolayer interferometry. Four new high-affinity laminin receptors were identified in the outer membrane; EstA, OprD, OprG and PA3923. Mutated bacteria devoid of these receptors adhered poorly to immobilized laminin. All bacterial receptors bound to the heparin-binding domains on LG4 and LG5 of the laminin alpha chain as assessed with truncated laminin fragments, transmission electron microscopy and inhibition by heparin. In conclusion, P. aeruginosa binds laminin via multiple surface receptors, and isolates from lungs of cystic fibrosis patients bound significantly more laminin compared to bacteria isolated from the skin and urine. Since laminin is abundant in both the lungs and skin, we suggest that laminin binding is an important mechanism in P. aeruginosa pathogenesis.

Peptidoglycan-Binding Anchor Is a Pseudomonas aeruginosa OmpA Family Lipoprotein With Importance for Outer Membrane Vesicles, Biofilms, and the Periplasmic Shape

“…The 6 × His-tagged proteins were expressed in E. coli BL21(DE3) (Novagen) after induction with 1 mM IPTG. Expressed proteins were purified on Ni-NTA resin columns (GE Healthcare, Chicago, IL) as previously described (Paulsson et al, 2019). Transcomplementation of the transposon mutant PW2884 was done by amplifying pba with 5 -ACTATTCGGATCCATTAAAGAGGAGAAATTACACATGAG CATCACGAGGAC-3 and 5 -AGTCATGACAAGCTTTCA TTCGCGCTTGATGAGGATTTCCA-3 , digested with BamHI and HindIII, and cloned into the complementation plasmid pUCP22Not.…”

“…The beads were incubated in PBS containing 1% milk and 0.05% Tween-20 (PMT) and transferred to 96-well V-bottom microtiter plates. His-tagged Pba or OprG as the control in PMT (Paulsson et al, 2019) was added to the beads and incubated for 1 h at 22 • C on a shaker (500 rpm). The beads were sedimented by gentle centrifugation (20 × g, 2 min), the excess liquid carefully removed, and then washed with PMT.…”

Pseudomonas aeruginosa prioritizes detoxification of hydrogen peroxide over nitric oxide

“…In particular, we investigated dual stress conditions in Pseudomonas aeruginosa , which differs significantly from E. coli despite both being Gram-negative bacteria. P. aeruginosa inhabits very different niches in the human body, such as the airways and skin, compared to E. coli , which thrives in the gastrointestinal system [ 13 , 14 ]. Genetically, P. aeruginosa and E. coli can harbor significantly different-sized genomes (e.g.…”

Heparan Sulfate Glycosaminoglycans: (Un)Expected Allies in Cancer Clinical Management

“…Among the pathogens that bind to host HS chains are parasites like Plasmodium falciparum [ 70 ]; bacteria, such as Escherichia coli [ 71 ], Pseudomonas aeruginosa [ 72 ], Borrelia burgdorferi [ 73 ] or Mycobacterium tuberculosis [ 74 ]; and many viruses, amongst which are found Human Papilloma virus, Herpes viruses and Human Immunodeficiency Virus-1 [ 75 , 76 ]. In addition, recently it was reported that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) entry in human cells is mediated through the binding to HS chains in a length and sequence-dependent manner [ 49 , 77 , 78 , 79 , 80 ].…”