Non-Specific Protein-Membrane Interactions: Adsorption and Fouling

Outer membrane vesicles (OMVs) act as carriers of bacterial products such as plasmids and resistance determinants, including metallo-β-lactamases. The lipidated, membrane-anchored metallo-β-lactamase NDM-1 can be detected in Gram-negative OMVs. The soluble domain of NDM-1 also forms electrostatic interactions with the membrane. Herein, we show that these interactions promote its packaging into OMVs produced by Escherichia coli . We report that favorable electrostatic protein-membrane interactions are also at work in the soluble enzyme IMP-1, while being absent in VIM-2. These interactions correlate with an enhanced incorporation of IMP-1 compared to VIM-2 into OMVs. Disruption of these interactions in NDM-1 and IMP-1 impairs their inclusion into vesicles, confirming their role in defining the protein cargo in OMVs. These results also indicate that packaging of metallo-β-lactamases into vesicles in their active form is a common phenomenon that involves cargo selection based on specific molecular interactions.

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Tug-of-war: molecular dynamometers against living cells for analyzing sub-piconewton interaction of specific protein with cell membrane

Chemical Science ◽

10.1039/d1sc03059k ◽

2021 ◽

Author(s):

Huipu Liu ◽

Yunlong Chen ◽

Jiawei Wang ◽

Yuanjiao Yang ◽

Huangxian Ju

Keyword(s):

Membrane Proteins ◽

Cell Membrane ◽

Specific Protein ◽

Living Cells ◽

Membrane Interactions ◽

Membrane Interaction ◽

Protein Membrane ◽

Functional Regulation ◽

Signal Transductions

Protein-membrane interactions play important roles in signal transductions and functional regulation of membrane proteins. Here, we design a molecular dynamometer (MDM) for analyzing protein-membrane interaction on living cells. The MDM...

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Specific protein-membrane interactions promote the export of metallo-β-lactamases via outer membrane vesicles

10.1101/2021.03.18.436108 ◽

2021 ◽

Author(s):

Carolina López ◽

Alessio Prunotto ◽

Guillermo Bahr ◽

Robert A. Bonomo ◽

Lisandro J. González ◽

...

Keyword(s):

Outer Membrane ◽

Electrostatic Interactions ◽

Active Form ◽

Membrane Vesicles ◽

Specific Protein ◽

Outer Membrane Vesicles ◽

Membrane Interactions ◽

Molecular Features ◽

Protein Membrane ◽

Enhanced Secretion

ABSTRACTOuter membrane vesicles (OMVs) act as carriers of resistance determinants such as metallo-β-lactamases. The metallo-β-lactamase NDM-1 is present in OMVs produced by Gram-negative bacteria since it is a lipidated, membrane-anchored protein. The soluble domain of NDM-1 also forms electrostatic interactions with the membrane. Herein, we show that these interactions promote its export into OMVs produced by Escherichia coli. We report that favorable electrostatic protein-membrane interactions are also at work in the soluble enzyme IMP-1, while being absent in VIM-2. These interactions correlate with an enhanced secretion into OMVs of IMP-1 compared to VIM-2. Disruption of these interactions in NDM-1 and IMP-1 impairs export into vesicles, confirming their role in defining the protein cargo in OMVs. These results also indicate that export of metallo-β-lactamases into vesicles in their active form is a common phenomenon that involves cargo selection based on molecular features.

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