Related Experiment Video

Updated: Oct 26, 2025

Directed Protein Packaging within Outer Membrane Vesicles from Escherichia coli: Design, Production and Purification
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Directed Protein Packaging within Outer Membrane Vesicles from Escherichia coli: Design, Production and Purification

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Specific Protein-Membrane Interactions Promote Packaging of Metallo-β-Lactamases into Outer Membrane Vesicles

Carolina López1, Alessio Prunotto2,3, Guillermo Bahr1,4

  • 1Instituto de Biología Molecular y Celular de Rosario (IBR, CONICET-UNR), Rosario, Argentina.

Antimicrobial Agents and Chemotherapy
|July 26, 2021
PubMed

Insights

Bacterial outer membrane vesicles (OMVs) package metallo-β-lactamases (e.g., NDM-1) via electrostatic interactions. These interactions determine which proteins are included in OMVs, impacting antibiotic resistance spread.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • Outer membrane vesicles (OMVs) are key mediators of bacterial intercellular communication and horizontal gene transfer.
  • Metallo-β-lactamases (MBLs) are crucial enzymes conferring antibiotic resistance, often found within OMVs.
  • The packaging mechanisms of MBLs into OMVs are not fully understood.

Purpose of the Study:

  • To investigate the role of protein-membrane electrostatic interactions in the packaging of metallo-β-lactamases into outer membrane vesicles.
  • To determine if these interactions influence the selection of specific MBLs for OMV cargo.

Main Methods:

  • Utilized *Escherichia coli* to produce OMVs.
  • Investigated electrostatic interactions between soluble enzyme domains and membranes for NDM-1, IMP-1, and VIM-2.
  • Assessed the impact of disrupting these interactions on MBL packaging into OMVs.

Main Results:

  • Lipidated NDM-1 and soluble NDM-1 domain engage in electrostatic interactions with the bacterial membrane, promoting OMV packaging.
  • Favorable electrostatic interactions were observed for IMP-1 but not VIM-2, correlating with differential OMV incorporation.
  • Disrupting electrostatic interactions significantly reduced NDM-1 and IMP-1 packaging into OMVs.

Conclusions:

  • Electrostatic protein-membrane interactions are critical determinants for selecting metallo-β-lactamase cargo within outer membrane vesicles.
  • This mechanism ensures the active form of MBLs is efficiently packaged, potentially facilitating the spread of antibiotic resistance.
  • OMV cargo selection is mediated by specific molecular interactions, offering insights into bacterial pathogenesis and resistance dissemination.
Keywords:
IMP-1NDM-1metallo-β-lactamasesouter membrane vesiclesprotein-membrane interactions

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