Two distinct anionic phospholipid-dependent events involved in SecA-mediated protein translocation

Koch, Sabrina; Exterkate, Marten; López Bautista, Cesar Augusto; Patro, Megha; Marrink, Siewert-Jan; Driessen, Arnold J. M.

doi:10.1016/j.bbamem.2019.183035

Title: Two distinct anionic phospholipid-dependent events involved in SecA-mediated protein translocation

Abstract

Protein translocation across the bacterial cytoplasmic membrane is an essential process catalyzed by the Sec translocase, which in its minimal form consists of the protein-conducting channel SecYEG, and the motor ATPase SecA. SecA binds via its positively charged N-terminus to membranes containing anionic phospholipids, leading to a lipid-bound intermediate. This interaction induces a conformational change in SecA, resulting in a high-affinity association with SecYEG, which begins protein translocation. In this work, we examined the effect of anionic lipids on the SecA-SecYEG interaction in more detail, and discovered a second, however unknown, anionic lipid-dependent event that stimulates protein translocation. Based on molecular dynamics simulations we identified an anionic lipid-enriched region in vicinity of the lateral gate of SecY. Here, the anionic lipid headgroup accesses the lateral gate, thereby stabilizing the pre-open state of the channel. The simulations suggest flip-flop movement of phospholipid along the lateral gate. Electrostatic contribution of the anionic phospholipids at the lateral gate may directly stabilize positively charged residues of the signal sequence of an incoming preprotein. Such a mechanism allows for the correct positioning of the entrant peptide, thereby providing a long-sought explanation for the role of anionic lipids in signal sequence folding during protein translocation.

Authors:

Koch, Sabrina ^[1]; Exterkate, Marten ^[1];

^[2]; Patro, Megha ^[1]; Marrink, Siewert-Jan ^[1]; Driessen, Arnold J. M. ^[1]

Univ. of Groningen (Netherlands)
Univ. of Groningen (Netherlands); Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

Publication Date:: Mon Aug 05 00:00:00 EDT 2019

Research Org.:: Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

Sponsoring Org.:: USDOE National Nuclear Security Administration (NNSA); Foundation for Fundamental Research on Matter (FOM)

OSTI Identifier:: 1558044

Report Number(s):: LA-UR-18-23216
Journal ID: ISSN 0005-2736

Grant/Contract Number:: 89233218CNA000001

Resource Type:: Accepted Manuscript

Journal Name:: Biochimica et Biophysica Acta. Biomembranes

Additional Journal Information:: Journal Volume: 1861; Journal Issue: 11; Journal ID: ISSN 0005-2736

Publisher:: Elsevier

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; Nanodiscs; SecYEG; Protein-lipid interactions; Protein translocation; Anionic phospholipids; Molecular dynamics

Citation Formats


                    Koch, Sabrina, Exterkate, Marten, López Bautista, Cesar Augusto, Patro, Megha, Marrink, Siewert-Jan, and Driessen, Arnold J. M. Two distinct anionic phospholipid-dependent events involved in SecA-mediated protein translocation.  United States: N. p., 2019. 
Web.  doi:10.1016/j.bbamem.2019.183035.

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                    Koch, Sabrina, Exterkate, Marten, López Bautista, Cesar Augusto, Patro, Megha, Marrink, Siewert-Jan, & Driessen, Arnold J. M. Two distinct anionic phospholipid-dependent events involved in SecA-mediated protein translocation.  United States.  https://doi.org/10.1016/j.bbamem.2019.183035

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                    Koch, Sabrina, Exterkate, Marten, López Bautista, Cesar Augusto, Patro, Megha, Marrink, Siewert-Jan, and Driessen, Arnold J. M. Mon .  
"Two distinct anionic phospholipid-dependent events involved in SecA-mediated protein translocation".  United States.  https://doi.org/10.1016/j.bbamem.2019.183035.  https://www.osti.gov/servlets/purl/1558044.

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@article{osti_1558044,

  title        = {Two distinct anionic phospholipid-dependent events involved in SecA-mediated protein translocation},

  author       = {Koch, Sabrina and Exterkate, Marten and López Bautista, Cesar Augusto and Patro, Megha and Marrink, Siewert-Jan and Driessen, Arnold J. M.},

  abstractNote = {Protein translocation across the bacterial cytoplasmic membrane is an essential process catalyzed by the Sec translocase, which in its minimal form consists of the protein-conducting channel SecYEG, and the motor ATPase SecA. SecA binds via its positively charged N-terminus to membranes containing anionic phospholipids, leading to a lipid-bound intermediate. This interaction induces a conformational change in SecA, resulting in a high-affinity association with SecYEG, which begins protein translocation. In this work, we examined the effect of anionic lipids on the SecA-SecYEG interaction in more detail, and discovered a second, however unknown, anionic lipid-dependent event that stimulates protein translocation. Based on molecular dynamics simulations we identified an anionic lipid-enriched region in vicinity of the lateral gate of SecY. Here, the anionic lipid headgroup accesses the lateral gate, thereby stabilizing the pre-open state of the channel. The simulations suggest flip-flop movement of phospholipid along the lateral gate. Electrostatic contribution of the anionic phospholipids at the lateral gate may directly stabilize positively charged residues of the signal sequence of an incoming preprotein. Such a mechanism allows for the correct positioning of the entrant peptide, thereby providing a long-sought explanation for the role of anionic lipids in signal sequence folding during protein translocation.},

  doi          = {10.1016/j.bbamem.2019.183035},

  journal      = {Biochimica et Biophysica Acta. Biomembranes},

  number       = 11,

  volume       = 1861,

  place        = {United States},

  year         = {Mon Aug 05 00:00:00 EDT 2019},

  month        = {Mon Aug 05 00:00:00 EDT 2019}

}

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https://doi.org/10.1016/j.bbamem.2019.183035

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Cited by: 8 works

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Figures / Tables:

Figure 1: Anionic lipid concentration dependency of the SecA-SecYEG interaction, translocation activity and ATPase activity. (a) Microscale Thermophoresis (MST) analysis of the binding of SecA to SecYEG reconstituted in nanodiscs harboring either CL (dark grey), PA (blue), PS (light grey), PG (black) or no anionic lipid (white). Fraction of boundmore »

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