Abstract

Significance The diverse environment of cellular membranes presents unique challenges in deciphering the roles that lipids play in modulating membrane protein structure and function. Here, we developed a native mass spectrometry approach to monitor binding of different lipid types to membrane proteins. We discovered that specific lipid−protein interactions can allosterically modulate the binding of lipids of different types. We also determined the structure of AmtB bound to cardiolipin, and mutation of residues involved in binding this lipid abolishes the observed allosteric effect. Our findings are of particular significance as they contribute to our general knowledge of how lipids modulate protein structure and function and how membrane proteins may recruit, through allostery, their own lipid microenvironment.