Lysine acetylation regulates the interaction between proteins and membranes

AbstractLysine acetylation regulates the function of soluble proteins in vivo, yet it remains largely unexplored whether lysine acetylation regulates membrane protein function. Here, we use bioinformatics, biophysical analysis of recombinant proteins, live-cell fluorescent imaging and genetic manipulation of Drosophila to explore lysine acetylation in peripheral membrane proteins. Analysis of 50 peripheral membrane proteins harboring BAR, PX, C2, or EHD membrane-binding domains reveals that lysine acetylation predominates in membrane-interaction regions. Acetylation and acetylation-mimicking mutations in three test proteins, amphiphysin, EHD2, and synaptotagmin1, strongly reduce membrane binding affinity, attenuate membrane remodeling in vitro and alter subcellular localization. This effect is likely due to the loss of positive charge, which weakens interactions with negatively charged membranes. In Drosophila, acetylation-mimicking mutations of amphiphysin cause severe disruption of T-tubule organization and yield a flightless phenotype. Our data provide mechanistic insights into how lysine acetylation regulates membrane protein function, potentially impacting a plethora of membrane-related processes.

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Mechanisms underlying drug-mediated regulation of membrane protein function

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.2113229118 ◽

2021 ◽

Vol 118 (46) ◽

pp. e2113229118

Author(s):

Radda Rusinova ◽

Changhao He ◽

Olaf S. Andersen

Keyword(s):

Membrane Proteins ◽

Membrane Protein ◽

Lipid Bilayer ◽

Conformational Changes ◽

Protein Function ◽

Nonspecific Binding ◽

Drug Induced ◽

Protein Conformational Changes ◽

Potassium Channel Kcsa ◽

Membrane Protein Function

The hydrophobic coupling between membrane proteins and their host lipid bilayer provides a mechanism by which bilayer-modifying drugs may alter protein function. Drug regulation of membrane protein function thus may be mediated by both direct interactions with the protein and drug-induced alterations of bilayer properties, in which the latter will alter the energetics of protein conformational changes. To tease apart these mechanisms, we examine how the prototypical, proton-gated bacterial potassium channel KcsA is regulated by bilayer-modifying drugs using a fluorescence-based approach to quantify changes in both KcsA function and lipid bilayer properties (using gramicidin channels as probes). All tested drugs inhibited KcsA activity, and the changes in the different gating steps varied with bilayer thickness, suggesting a coupling to the bilayer. Examining the correlations between changes in KcsA gating steps and bilayer properties reveals that drug-induced regulation of membrane protein function indeed involves bilayer-mediated mechanisms. Both direct, either specific or nonspecific, binding and bilayer-mediated mechanisms therefore are likely to be important whenever there is overlap between the concentration ranges at which a drug alters membrane protein function and bilayer properties. Because changes in bilayer properties will impact many diverse membrane proteins, they may cause indiscriminate changes in protein function.

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Synthesis of Extended, Self-Assembled Biological Membranes containing Membrane Proteins from Gas Phase

10.1101/661215 ◽

2019 ◽

Cited By ~ 1

Author(s):

Matthias Wilm

Keyword(s):

Membrane Proteins ◽

Membrane Protein ◽

Self Assembly ◽

Protein Function ◽

Lipid Membranes ◽

Protein Complexes ◽

Biological Membranes ◽

The Self ◽

In Vitro Systems

1.AbstractMembrane proteins carry out a wide variety of biological functions. The reproduction of specific properties that have evolved over millions of years of biological membranes in a technically controlled environment is of significant interest. Here a method is presented that allows the self-assembly of a macroscopically large, freely transportable membrane with Outer membrane porin G from Escherichia Coli. The technique does not use protein specific characteristics and therefore, could represent a method for the generation of extended layers of membranes with arbitrary membrane protein content. Such in-vitro systems are relevant in the study of membrane-protein function and structure and the self-assembly of membrane-based protein complexes. They might become important for the incorporation of the lipid-membranes in technological devices.

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Phosphoinositide Control of Membrane Protein Function: A Frontier Led by Studies on Ion Channels

Annual Review of Physiology ◽

10.1146/annurev-physiol-021113-170358 ◽

2015 ◽

Vol 77 (1) ◽

pp. 81-104 ◽

Cited By ~ 51

Author(s):

Diomedes E. Logothetis ◽

Vasileios I. Petrou ◽

Miao Zhang ◽

Rahul Mahajan ◽

Xuan-Yu Meng ◽

...

Keyword(s):

Ion Channels ◽

Membrane Protein ◽

Protein Function ◽

Membrane Protein Function

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Sirtuin-dependent reversible lysine acetylation controls the activity of acetyl-Coenzyme A synthetase in Campylobacter jejuni

Journal of Bacteriology ◽

10.1128/jb.00333-21 ◽

2021 ◽

Author(s):

Victoria L. Jeter ◽

Jorge C. Escalante-Semerena

Keyword(s):

Campylobacter Jejuni ◽

Posttranslational Modifications ◽

Active Site ◽

Protein Function ◽

Metabolic Enzyme ◽

Lysine Acetylation ◽

Class Iii ◽

Acetyl Coa

Posttranslational modifications are mechanisms for rapid control of protein function used by cells from all domains of life. Acetylation of the epsilon amino group ( N ε ) of an active-site lysine of the AMP-forming acetyl-CoA synthetase (Acs) enzyme is the paradigm for the posttranslational control of the activity of metabolic enzymes. In bacteria, the alluded active-site lysine of Acs enzymes can be modified by a number of different GCN5-type N -acetyltransferases (GNATs). Acs activity is lost as a result of acetylation, and restored by deacetylation. Using a heterologous host, we show that Campylobacter jejuni NCTC11168 synthesizes enzymes that control Acs function by reversible lysine acetylation (RLA). This work validates the function of gene products encoded by the cj1537c , cj1715, and cj1050c loci, namely the AMP-forming acetate:CoA ligase ( Cj Acs), a type IV GCN5-type lysine acetyltransferase (GNAT, hereafter Cj LatA), and a NAD + -dependent (class III) sirtuin deacylase ( Cj CobB), respectively. To our knowledge, these are the first in vivo and in vitro data on C. jejuni enzymes that control the activity of Cj Acs. IMPORTANCE This work is important because it provides the experimental evidence needed to support the assignment of function to three key enzymes, two of which control the reversible posttranslational modification of an active-site lysyl residue of the central metabolic enzyme acetyl-CoA synthetase ( Cj Acs). We can now generate Campylobacter jejuni mutant strains defective in these functions, so we can establish the conditions in which this mode of regulation of Cj Acs is triggered in this bacterium. Such knowledge may provide new therapeutic strategies for the control of this pathogen.

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