FA Sliding As the Mechanism for the ANT1-Mediated Fatty Acid Anion Transport in Lipid Bilayers

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2023 Sep 28

PMID 37762012

Authors

Jurgen Kreiter

Sanja Skulj

Zlatko Brkljaca

Sarah Bardakji

Mario Vazdar

Elena E Pohl

Affiliations

Soon will be listed here.

Abstract

Mitochondrial adenine nucleotide translocase (ANT) exchanges ADP for ATP to maintain energy production in the cell. Its protonophoric function in the presence of long-chain fatty acids (FA) is also recognized. Our previous results imply that proton/FA transport can be best described with the FA cycling model, in which protonated FA transports the proton to the mitochondrial matrix. The mechanism by which ANT1 transports FA anions back to the intermembrane space remains unclear. Using a combined approach involving measurements of the current through the planar lipid bilayers reconstituted with ANT1, site-directed mutagenesis and molecular dynamics simulations, we show that the FA anion is first attracted by positively charged arginines or lysines on the matrix side of ANT1 before moving along the positively charged protein-lipid interface and binding to R79, where it is protonated. We show that R79 is also critical for the competitive binding of ANT1 substrates (ADP and ATP) and inhibitors (carboxyatractyloside and bongkrekic acid). The binding sites are well conserved in mitochondrial SLC25 members, suggesting a general mechanism for transporting FA anions across the inner mitochondrial membrane.

Citing Articles

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References

Mjos O . Influence of free fatty acids on myocardial oxygen consumption and ischemic injury. Am J Cardiol. 1981; 48(2):361-5. DOI: 10.1016/0002-9149(81)90621-4. View

Wojtczak L, Wieckowski M, Schonfeld P . Protonophoric activity of fatty acid analogs and derivatives in the inner mitochondrial membrane: a further argument for the fatty acid cycling model. Arch Biochem Biophys. 1998; 357(1):76-84. DOI: 10.1006/abbi.1998.0777. View

Palmieri F . The mitochondrial transporter family SLC25: identification, properties and physiopathology. Mol Aspects Med. 2012; 34(2-3):465-84. DOI: 10.1016/j.mam.2012.05.005. View

Ardalan A, Sowlati-Hashjin S, Uwumarenogie S, Fish M, Mitchell J, Karttunen M . Functional Oligomeric Forms of Uncoupling Protein 2: Strong Evidence for Asymmetry in Protein and Lipid Bilayer Systems. J Phys Chem B. 2020; 125(1):169-183. DOI: 10.1021/acs.jpcb.0c09422. View

Kamp F, Zakim D, Zhang F, Noy N, Hamilton J . Fatty acid flip-flop in phospholipid bilayers is extremely fast. Biochemistry. 1995; 34(37):11928-37. DOI: 10.1021/bi00037a034. View

Mifsud J, Ravaud S, Krammer E, Chipot C, Kunji E, Pebay-Peyroula E . The substrate specificity of the human ADP/ATP carrier AAC1. Mol Membr Biol. 2012; 30(2):160-8. DOI: 10.3109/09687688.2012.745175. View

Ardalan A, Sowlati-Hashjin S, Oduwoye H, Uwumarenogie S, Karttunen M, Smith M . Biphasic Proton Transport Mechanism for Uncoupling Proteins. J Phys Chem B. 2021; 125(32):9130-9144. DOI: 10.1021/acs.jpcb.1c04766. View

Beck V, Jaburek M, Demina T, Rupprecht A, Porter R, Jezek P . Polyunsaturated fatty acids activate human uncoupling proteins 1 and 2 in planar lipid bilayers. FASEB J. 2007; 21(4):1137-44. DOI: 10.1096/fj.06-7489com. View

Samartsev V, Smirnov A, Zeldi I, Markova O, Mokhova E, Skulachev V . Involvement of aspartate/glutamate antiporter in fatty acid-induced uncoupling of liver mitochondria. Biochim Biophys Acta. 1997; 1319(2-3):251-7. DOI: 10.1016/s0005-2728(96)00166-1. View

10.

Krauss S, Zhang C, Lowell B . The mitochondrial uncoupling-protein homologues. Nat Rev Mol Cell Biol. 2005; 6(3):248-61. DOI: 10.1038/nrm1592. View

11.

Palmieri L, Agrimi G, Runswick M, Fearnley I, Palmieri F, Walker J . Identification in Saccharomyces cerevisiae of two isoforms of a novel mitochondrial transporter for 2-oxoadipate and 2-oxoglutarate. J Biol Chem. 2000; 276(3):1916-22. DOI: 10.1074/jbc.M004332200. View

12.

Mavridou V, King M, Tavoulari S, Ruprecht J, Palmer S, Kunji E . Substrate binding in the mitochondrial ADP/ATP carrier is a step-wise process guiding the structural changes in the transport cycle. Nat Commun. 2022; 13(1):3585. PMC: 9226169. DOI: 10.1038/s41467-022-31366-5. View

13.

King M, Kerr M, Crichton P, Springett R, Kunji E . Formation of a cytoplasmic salt bridge network in the matrix state is a fundamental step in the transport mechanism of the mitochondrial ADP/ATP carrier. Biochim Biophys Acta. 2015; 1857(1):14-22. PMC: 4674015. DOI: 10.1016/j.bbabio.2015.09.013. View

14.

Beck V, Jaburek M, Breen E, Porter R, Jezek P, Pohl E . A new automated technique for the reconstitution of hydrophobic proteins into planar bilayer membranes. Studies of human recombinant uncoupling protein 1. Biochim Biophys Acta. 2006; 1757(5-6):474-9. DOI: 10.1016/j.bbabio.2006.03.006. View

15.

LaNoue K, Mizani S, Klingenberg M . Electrical imbalance of adenine nucleotide transport across the mitochondrial membrane. J Biol Chem. 1978; 253(1):191-8. View

16.

Chipot C, Dehez F, Schnell J, Zitzmann N, Pebay-Peyroula E, Catoire L . Perturbations of Native Membrane Protein Structure in Alkyl Phosphocholine Detergents: A Critical Assessment of NMR and Biophysical Studies. Chem Rev. 2018; 118(7):3559-3607. PMC: 5896743. DOI: 10.1021/acs.chemrev.7b00570. View

17.

Bertholet A, Kirichok Y . The Mechanism FA-Dependent H Transport by UCP1. Handb Exp Pharmacol. 2018; 251:143-159. DOI: 10.1007/164_2018_138. View

18.

Humphrey W, Dalke A, Schulten K . VMD: visual molecular dynamics. J Mol Graph. 1996; 14(1):33-8, 27-8. DOI: 10.1016/0263-7855(96)00018-5. View

19.

Skulachev V . Fatty acid circuit as a physiological mechanism of uncoupling of oxidative phosphorylation. FEBS Lett. 1991; 294(3):158-62. DOI: 10.1016/0014-5793(91)80658-p. View

20.

Jo S, Kim T, Im W . Automated builder and database of protein/membrane complexes for molecular dynamics simulations. PLoS One. 2007; 2(9):e880. PMC: 1963319. DOI: 10.1371/journal.pone.0000880. View