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Title: Ladderane phospholipids form a densely packed membrane with normal hydrazine and anomalously low proton/hydroxide permeability

Abstract

Ladderane lipids are unique to anaerobic ammonium-oxidizing (anammox) bacteria and are enriched in the membrane of the anammoxosome, an organelle thought to compartmentalize the anammox process, which involves the toxic intermediate hydrazine (N 2H 4). Due to the slow growth rate of anammox bacteria and difficulty of isolating pure ladderane lipids, experimental evidence of the biological function of ladderanes is lacking. We have synthesized two natural and one unnatural ladderane phosphatidylcholine lipids and compared their thermotropic properties in self-assembled bilayers to distinguish between [3]- and [5]-ladderane function. We then developed a hydrazine transmembrane diffusion assay using a water-soluble derivative of a hydrazine sensor and determined that ladderane membranes are as permeable to hydrazine as straight-chain lipid bilayers. However, pH equilibration across ladderane membranes occurs 5–10 times more slowly than across straight-chain lipid membranes. Langmuir monolayer analysis and the rates of fluorescence recovery after photobleaching suggest that dense ladderane packing may preclude formation of proton/hydroxide-conducting water wires. These data support the hypothesis that ladderanes prevent the breakdown of the proton motive force rather than blocking hydrazine transmembrane diffusion in anammox bacteria.

Authors:
 [1];  [1];  [1];  [1];  [2];  [1];  [1]
  1. Stanford Univ., CA (United States). Dept. of Chemistry
  2. SLAC National Accelerator Lab., Menlo Park, CA (United States). Stanford Synchrotron Radiation Lightsource (SSRL)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); National Institutes of Health (NIH); National Science Foundation (NSF)
OSTI Identifier:
1476321
Grant/Contract Number:  
AC02-76SF00515; GM069630; GM118044; P41GM103393; ECCS-1542152
Resource Type:
Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 115; Journal Issue: 37; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; ladderane; anammox; lipid bilayer; proton permeability; membrane structure

Citation Formats

Moss, Frank R., Shuken, Steven R., Mercer, Jaron A. M., Cohen, Carolyn M., Weiss, Thomas M., Boxer, Steven G., and Burns, Noah Z. Ladderane phospholipids form a densely packed membrane with normal hydrazine and anomalously low proton/hydroxide permeability. United States: N. p., 2018. Web. doi:10.1073/pnas.1810706115.
Moss, Frank R., Shuken, Steven R., Mercer, Jaron A. M., Cohen, Carolyn M., Weiss, Thomas M., Boxer, Steven G., & Burns, Noah Z. Ladderane phospholipids form a densely packed membrane with normal hydrazine and anomalously low proton/hydroxide permeability. United States. doi:10.1073/pnas.1810706115.
Moss, Frank R., Shuken, Steven R., Mercer, Jaron A. M., Cohen, Carolyn M., Weiss, Thomas M., Boxer, Steven G., and Burns, Noah Z. Mon . "Ladderane phospholipids form a densely packed membrane with normal hydrazine and anomalously low proton/hydroxide permeability". United States. doi:10.1073/pnas.1810706115. https://www.osti.gov/servlets/purl/1476321.
@article{osti_1476321,
title = {Ladderane phospholipids form a densely packed membrane with normal hydrazine and anomalously low proton/hydroxide permeability},
author = {Moss, Frank R. and Shuken, Steven R. and Mercer, Jaron A. M. and Cohen, Carolyn M. and Weiss, Thomas M. and Boxer, Steven G. and Burns, Noah Z.},
abstractNote = {Ladderane lipids are unique to anaerobic ammonium-oxidizing (anammox) bacteria and are enriched in the membrane of the anammoxosome, an organelle thought to compartmentalize the anammox process, which involves the toxic intermediate hydrazine (N2H4). Due to the slow growth rate of anammox bacteria and difficulty of isolating pure ladderane lipids, experimental evidence of the biological function of ladderanes is lacking. We have synthesized two natural and one unnatural ladderane phosphatidylcholine lipids and compared their thermotropic properties in self-assembled bilayers to distinguish between [3]- and [5]-ladderane function. We then developed a hydrazine transmembrane diffusion assay using a water-soluble derivative of a hydrazine sensor and determined that ladderane membranes are as permeable to hydrazine as straight-chain lipid bilayers. However, pH equilibration across ladderane membranes occurs 5–10 times more slowly than across straight-chain lipid membranes. Langmuir monolayer analysis and the rates of fluorescence recovery after photobleaching suggest that dense ladderane packing may preclude formation of proton/hydroxide-conducting water wires. These data support the hypothesis that ladderanes prevent the breakdown of the proton motive force rather than blocking hydrazine transmembrane diffusion in anammox bacteria.},
doi = {10.1073/pnas.1810706115},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 37,
volume = 115,
place = {United States},
year = {2018},
month = {8}
}

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Works referenced in this record:

How to make a living from anaerobic ammonium oxidation
journal, May 2013

  • Kartal, Boran; de Almeida, Naomi M.; Maalcke, Wouter J.
  • FEMS Microbiology Reviews, Vol. 37, Issue 3
  • DOI: 10.1111/1574-6976.12014

Formation and properties of thin-walled phospholipid vesicles
journal, February 1969

  • Reeves, John P.; Dowben, Robert M.
  • Journal of Cellular Physiology, Vol. 73, Issue 1
  • DOI: 10.1002/jcp.1040730108

Giant Vesicles: Preparations and Applications
journal, March 2010


Model Systems, Lipid Rafts, and Cell Membranes
journal, June 2004


Attack of radicals and protons on ladderane lipids: quantum chemical calculations and biological implications
journal, January 2012

  • Nouri, Dustin H.; Tantillo, Dean J.
  • Organic & Biomolecular Chemistry, Vol. 10, Issue 29
  • DOI: 10.1039/c2ob25717c

Insights into the role of cyclic ladderane lipids in bacteria from computer simulations
journal, July 2014


Molecular mechanism of anaerobic ammonium oxidation
journal, October 2011

  • Kartal, Boran; Maalcke, Wouter J.; de Almeida, Naomi M.
  • Nature, Vol. 479, Issue 7371, p. 127-130
  • DOI: 10.1038/nature10453

Molecular Mechanisms of Water and Solute Transport across Archaebacterial Lipid Membranes
journal, May 2001

  • Mathai, John C.; Sprott, G. Dennis; Zeidel, Mark L.
  • Journal of Biological Chemistry, Vol. 276, Issue 29
  • DOI: 10.1074/jbc.M103265200

Biophysical properties of membrane lipids of anammox bacteria: II. Impact of temperature and bacteriohopanoids
journal, July 2009

  • Boumann, Henry A.; Stroeve, Pieter; Longo, Marjorie L.
  • Biochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 1788, Issue 7
  • DOI: 10.1016/j.bbamem.2009.04.005

A mixed ladderane/n-alkyl glycerol diether membrane lipid in an anaerobic ammonium-oxidizing bacterium
journal, January 2004

  • Sinninghe Damsté, Jaap S.; Rijpstra, W. Irene C.; Strous, Marc
  • Chem. Commun., Issue 22
  • DOI: 10.1039/B409806D

Permeability of small nonelectrolytes through lipid bilayer membranes
journal, October 1986

  • Walter, Anne; Gutknecht, John
  • The Journal of Membrane Biology, Vol. 90, Issue 3
  • DOI: 10.1007/BF01870127

Biophysical properties of membrane lipids of anammox bacteria: I. Ladderane phospholipids form highly organized fluid membranes
journal, July 2009

  • Boumann, Henry A.; Longo, Marjorie L.; Stroeve, Pieter
  • Biochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 1788, Issue 7
  • DOI: 10.1016/j.bbamem.2009.04.008

Proton flux mechanisms in model and biological membranes
journal, February 1989

  • Deamer, David W.; Nichols, J. Wylie
  • The Journal of Membrane Biology, Vol. 107, Issue 2
  • DOI: 10.1007/BF01871715

The sequencing batch reactor as a powerful tool for the study of slowly growing anaerobic ammonium-oxidizing microorganisms
journal, November 1998

  • Strous, M.; Heijnen, J. J.; Kuenen, J. G.
  • Applied Microbiology and Biotechnology, Vol. 50, Issue 5
  • DOI: 10.1007/s002530051340

Anammox bacteria: from discovery to application
journal, April 2008


Permeation of protons, potassium ions, and small polar molecules through phospholipid bilayers as a function of membrane thickness
journal, January 1996


A new fluorescent and colorimetric sensor for hydrazine and its application in biological systems
journal, January 2014

  • Sun, Mingda; Guo, Jing; Yang, Qingbiao
  • J. Mater. Chem. B, Vol. 2, Issue 13
  • DOI: 10.1039/C3TB21753A

Ladderane phospholipids in anammox bacteria comprise phosphocholine and phosphoethanolamine headgroups
journal, May 2006


Membrane lipids: where they are and how they behave
journal, February 2008

  • van Meer, Gerrit; Voelker, Dennis R.; Feigenson, Gerald W.
  • Nature Reviews Molecular Cell Biology, Vol. 9, Issue 2
  • DOI: 10.1038/nrm2330

Protons May Leak through Pure Lipid Bilayers via a Concerted Mechanism
journal, May 2005


They Came From the Deep: Syntheses, Applications, and Biology of Ladderanes
journal, November 2006


Structural determinants of protein partitioning into ordered membrane domains and lipid rafts
journal, November 2015


Structural Determinants of Water Permeability through the Lipid Membrane
journal, December 2007

  • Mathai, John C.; Tristram-Nagle, Stephanie; Nagle, John F.
  • The Journal of General Physiology, Vol. 131, Issue 1
  • DOI: 10.1085/jgp.200709848

Modulation of Plasma Membrane Ca 2+ -ATPase by Neutral Phospholipids : EFFECT OF THE MICELLE-VESICLE TRANSITION AND THE BILAYER THICKNESS
journal, January 2015

  • Pignataro, María Florencia; Dodes-Traian, Martín M.; González-Flecha, F. Luis
  • Journal of Biological Chemistry, Vol. 290, Issue 10
  • DOI: 10.1074/jbc.M114.585828

Preparation of giant liposomes in physiological conditions and their characterization under an optical microscope
journal, December 1996


An intracellular pH gradient in the anammox bacterium Kuenenia stuttgartiensis as evaluated by 31P NMR
journal, October 2009

  • van der Star, Wouter R. L.; Dijkema, Cor; de Waard, Pieter
  • Applied Microbiology and Biotechnology, Vol. 86, Issue 1
  • DOI: 10.1007/s00253-009-2309-9

Structural identification of ladderane and other membrane lipids of planctomycetes capable of anaerobic ammonium oxidation (anammox): Membrane lipids of anammox bacteria
journal, August 2005


The inner workings of the hydrazine synthase multiprotein complex
journal, October 2015

  • Dietl, Andreas; Ferousi, Christina; Maalcke, Wouter J.
  • Nature, Vol. 527, Issue 7578
  • DOI: 10.1038/nature15517

Proton transport across transient single-file water pores in a lipid membrane studied by molecular dynamics simulations
journal, August 1996


Cation permeability induced by valinomycin, gramicidin D and amphotericin B in large lipidic unilamellar vesicles studied by 31P-NMR
journal, August 1985

  • Herv�, M.; Cybulska, B.; Gary-Bobo, C. M.
  • European Biophysics Journal, Vol. 12, Issue 3
  • DOI: 10.1007/BF00254070

The anammoxosome: an intracytoplasmic compartment in anammox bacteria
journal, April 2004

  • Niftrik, Laura A.; Fuerst, John A.; Damsté, Jaap S. Sinninghe
  • FEMS Microbiology Letters, Vol. 233, Issue 1
  • DOI: 10.1016/j.femsle.2004.01.044

Cyclohexane Rings Reduce Membrane Permeability to Small Ions in Archaea-Inspired Tetraether Lipids
journal, December 2015

  • Koyanagi, Takaoki; Leriche, Geoffray; Onofrei, David
  • Angewandte Chemie International Edition, Vol. 55, Issue 5
  • DOI: 10.1002/anie.201510445

Diffusion of univalent ions across the lamellae of swollen phospholipids
journal, August 1965


Nature Utilizes Unusual High London Dispersion Interactions for Compact Membranes Composed of Molecular Ladders
journal, February 2014

  • Wagner, J. Philipp; Schreiner, Peter R.
  • Journal of Chemical Theory and Computation, Vol. 10, Issue 3
  • DOI: 10.1021/ct5000499

Separation of Liquid Phases in Giant Vesicles of Ternary Mixtures of Phospholipids and Cholesterol
journal, November 2003


Linearly concatenated cyclobutane lipids form a dense bacterial membrane
journal, October 2002

  • Sinninghe Damsté, Jaap S.; Strous, Marc; Rijpstra, W. Irene C.
  • Nature, Vol. 419, Issue 6908
  • DOI: 10.1038/nature01128

Ladderane lipid distribution in four genera of anammox bacteria
journal, April 2008

  • Rattray, Jayne E.; van de Vossenberg, Jack; Hopmans, Ellen C.
  • Archives of Microbiology, Vol. 190, Issue 1
  • DOI: 10.1007/s00203-008-0364-8

Choose Your Label Wisely: Water-Soluble Fluorophores Often Interact with Lipid Bilayers
journal, February 2014


The membrane bioreactor: A novel tool to grow anammox bacteria as free cells
journal, October 2008

  • van der Star, Wouter R. L.; Miclea, Andreea I.; van Dongen, Udo G. J. M.
  • Biotechnology and Bioengineering, Vol. 101, Issue 2
  • DOI: 10.1002/bit.21891

Widespread distribution of encapsulin nanocompartments reveals functional diversity
journal, March 2017


Chemical Synthesis and Self-Assembly of a Ladderane Phospholipid
journal, November 2016

  • Mercer, Jaron A. M.; Cohen, Carolyn M.; Shuken, Steven R.
  • Journal of the American Chemical Society, Vol. 138, Issue 49
  • DOI: 10.1021/jacs.6b10706

Biochemistry and molecular biology of anammox bacteria
journal, June 2009

  • Jetten, Mike S. M.; Niftrik, Laura van; Strous, Marc
  • Critical Reviews in Biochemistry and Molecular Biology, Vol. 44, Issue 2-3
  • DOI: 10.1080/10409230902722783

    Works referencing / citing this record:

    Lessons in Strain and Stability: Enantioselective Synthesis of (+)-[5]-Ladderanoic Acid
    journal, November 2019

    • Hancock, Erin N.; Kuker, Erin L.; Tantillo, Dean J.
    • Angewandte Chemie International Edition, Vol. 59, Issue 1
    • DOI: 10.1002/anie.201910901

    Lessons in Strain and Stability: Enantioselective Synthesis of (+)-[5]-Ladderanoic Acid
    journal, November 2019

    • Hancock, Erin N.; Kuker, Erin L.; Tantillo, Dean J.
    • Angewandte Chemie International Edition, Vol. 59, Issue 1
    • DOI: 10.1002/anie.201910901