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Cited by 81Extracellular vesicles in diseaseIon Transport and Channel RegulationCellular transport and secretion

Exosomal GAPDH from Proximal Tubule Cells Regulate ENaC Activity

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Jella, Kishore Kumar, Yu, Ling, Yue, Qiang, Friedman, Daniel, Duke, Billie J., Alli, Abdel A.·2016·PLOS ONE
DOI 10.1371/journal.pone.0165763PMC PMC5089749
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Abstract

Exosomes are nanometer-scale, cell-derived vesicles that contain various molecules including nucleic acids, proteins, and lipids. These vesicles can release their cargo into adjacent or distant cells and mediate intercellular communication and cellular function. Here we examined the regulation of epithelial sodium channels in mpkCCD cells and distal tubule Xenopus 2F3 cells by exosomes isolated from proximal tubule LLC-PK1 cells. Cultured mpkCCD cells were stained with CTX coupled to a green fluorophore in order to label the cell membranes and freshly isolated exosomes from LLC-PK1 cells were labeled with the red lipophilic dye PKH26 in order to visualize uptake of exosomes into the cells. Single-channel patch clamp recordings showed the open probability of ENaC in Xenopus 2F3 cells and in freshly isolated split-open tubules decreased in response to exogenous application of exosomes derived from LLC-PK1 proximal tubule cells. Active GAPDH was identified within exosomes derived from proximal tubule LLC-PK1 cells. The effect on ENaC activity in Xenopus 2F3 cells was blunted after application of exosomes transfected with the GAPDH inhibitor heptelidic acid. Also, we show GAPDH and ENaC subunits associate in mpkCCD cells. These studies examine a potential role for exosomes in the regulation of ENaC activity and examine a possible mechanism for communication from proximal tubule cells to distal tubule and collecting duct cells.

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  • Some studies have reported GAPDH within the EV lumen, while others have identified it on the EV outer surface.

    “Some studies have reported the presence of GAPDH within the EV lumen, but others have identified it on the EV’s outer surface33,62.”

    From GAPDH controls extracellular vesicle biogenesis and enhances the therapeutic potential of EV mediated siRNA delivery to the brain (2021) · cites this paper as [62]

    The Results section ('Exosomes from LLC-PK1 cells contain active GAPDH'; page unavailable) reports GAPDH activity after exosomes were lysed and suggests that GAPDH may remain active inside them. It does not identify GAPDH on the EV outer surface, and the wording about its internal location is tentative. (LLM, conf=0.85)

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  1. [1]Vallon V, Rieg T. Regulation of renal NaCl and water transport by the ATP/UTP/P2Y2 receptor system. Am J Physiol Renal Physiol. 2011; 301(3): F463– 75. 10.1152/ajprenal.00236.2011 21715471 PMC3174559 · DOI
  2. [2]Gildea JJ, Seaton JE, Victor KG, Reyes CM, Bigler Wang D, Pettigrew AC, et al Exosomal transfer from human renal proximal tubule cells to distal tubule and collecting duct cells. Clin Biochem. 2014; 47(15): 89– 94. 10.1016/j.clinbiochem.2014.06.018. 24976626 PMC5384615 · DOI
  3. [3]Valadi H, Ekstrom K, Bossios A, Sjostrand M, Lee JJ, Lotvall JO. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol. 2007; 9(6): 654– 9. 10.1038/ncb1596. 17486113 · DOI · Paper page
  4. [4]Raposo G, Stoorvogel W. Extracellular vesicles: exosomes, microvesicles, and friends. J Cell Biol. 2013; 200(4): 373– 83. 10.1083/jcb.201211138 23420871 PMC3575529 · DOI · Paper page
  5. [5]van Balkom BW, Pisitkun T, Verhaar MC, Knepper MA. Exosomes and the kidney: prospects for diagnosis and therapy of renal diseases. Kidney Int. 2011; 80(11): 1138– 45. 10.1038/ki.2011.292 21881557 PMC3412193 · DOI
  6. [6]Jovov B, Tousson A, Ji HL, Keeton D, Shlyonsky V, Ripoll PJ, et al Regulation of epithelial Na(+) channels by actin in planar lipid bilayers and in the Xenopus oocyte expression system. J Biol Chem. 1999; 274(53): 37845– 54.. 10608849
  7. [7]Malik B, Price SR, Mitch WE, Yue Q, Eaton DC. Regulation of epithelial sodium channels by the ubiquitin-proteasome proteolytic pathway. Am J Physiol Renal Physiol. 2006; 290(6): F1285– 94. 10.1152/ajprenal.00432.2005. 16682484 · DOI
  8. [8]Alli AA, Song JZ, Al-Khalili O, Bao HF, Ma HP, Alli AA, et al Cathepsin B is secreted apically from Xenopus 2F3 cells and cleaves the epithelial sodium channel (ENaC) to increase its activity. J Biol Chem. 2012; 287(36): 30073– 83. 10.1074/jbc.M111.338574 22782900 PMC3436264 · DOI
  9. [9]Bruns JB, Carattino MD, Sheng S, Maarouf AB, Weisz OA, Pilewski JM, et al Epithelial Na+ channels are fully activated by furin- and prostasin-dependent release of an inhibitory peptide from the gamma-subunit. J Biol Chem. 2007; 282(9): 6153– 60. 10.1074/jbc.M610636200. 17199078 · DOI
  10. [10]Caldwell RA, Boucher RC, Stutts MJ. Neutrophil elastase activates near-silent epithelial Na+ channels and increases airway epithelial Na+ transport. Am J Physiol Lung Cell Mol Physiol. 2005; 288(5): L813– 9. 10.1152/ajplung.00435.2004. 15640288 · DOI
  11. [11]Haerteis S, Krappitz M, Bertog M, Krappitz A, Baraznenok V, Henderson I, et al Proteolytic activation of the epithelial sodium channel (ENaC) by the cysteine protease cathepsin-S. Pflugers Arch. 2012; 464(4): 353– 65. 10.1007/s00424-012-1138-3 22864553 PMC3448907 · DOI
  12. [12]Hughey RP, Bruns JB, Kinlough CL, Harkleroad KL, Tong Q, Carattino MD, et al Epithelial sodium channels are activated by furin-dependent proteolysis. J Biol Chem. 2004; 279(18): 18111– 4. 10.1074/jbc.C400080200. 15007080 · DOI
  13. [13]Kleyman TR, Carattino MD, Hughey RP. ENaC at the cutting edge: regulation of epithelial sodium channels by proteases. J Biol Chem. 2009; 284(31): 20447– 51. 10.1074/jbc.R800083200 19401469 PMC2742807 · DOI
  14. [14]Passero CJ, Mueller GM, Rondon-Berrios H, Tofovic SP, Hughey RP, Kleyman TR. Plasmin activates epithelial Na+ channels by cleaving the gamma subunit. J Biol Chem. 2008; 283(52): 36586– 91. 10.1074/jbc.M805676200 18981180 PMC2605981 · DOI
  15. [15]Patel AB, Chao J, Palmer LG. Tissue kallikrein activation of the epithelial Na channel. Am J Physiol Renal Physiol. 2012; 303(4): F540– 50. 10.1152/ajprenal.00133.2012 22622459 PMC3423117 · DOI
  16. [16]Ma HP, Saxena S, Warnock DG. Anionic phospholipids regulate native and expressed epithelial sodium channel (ENaC). J Biol Chem. 2002; 277(10): 7641– 4. 10.1074/jbc.C100737200. 11809744 · DOI
  17. [17]Pochynyuk O, Bugaj V, Vandewalle A, Stockand JD. Purinergic control of apical plasma membrane PI(4,5)P2 levels sets ENaC activity in principal cells. Am J Physiol Renal Physiol. 2008; 294(1): F38– 46. 10.1152/ajprenal.00403.2007. 17913833 · DOI
  18. [18]Sun Y, Zhang JN, Zhao D, Wang QS, Gu YC, Ma HP, et al Role of the epithelial sodium channel in salt-sensitive hypertension. Acta Pharmacol Sin. 2011; 32(6): 789– 97. 10.1038/aps.2011.72 21623391 PMC4009973 · DOI
  19. [19]Alli AA, Bao HF, Liu BC, Yu L, Aldrugh S, Montgomery DS, et al Calmodulin and CaMKII modulate ENaC activity by regulating the association of MARCKS and the cytoskeleton with the apical membrane. Am J Physiol Renal Physiol. 2015; 309(5): F456– 63. 10.1152/ajprenal.00631.2014 26136560 PMC4556887 · DOI
  20. [20]Yue G, Malik B, Yue G, Eaton DC. Phosphatidylinositol 4,5-bisphosphate (PIP2) stimulates epithelial sodium channel activity in A6 cells. J Biol Chem. 2002; 277(14): 11965– 9. 10.1074/jbc.M108951200. 11812779 · DOI
  21. [21]Ma HP, Eaton DC. Acute regulation of epithelial sodium channel by anionic phospholipids. J Am Soc Nephrol. 2005; 16(11): 3182– 7. 10.1681/ASN.2005040434. 16192420 · DOI
  22. [22]Ilatovskaya DV, Pavlov TS, Levchenko V, Negulyaev YA, Staruschenko A. Cortical actin binding protein cortactin mediates ENaC activity via Arp2/3 complex. FASEB J. 2011; 25(8): 2688– 99. 10.1096/fj.10-167262. 21536685 · DOI
  23. [23]Reifenberger MS, Yu L, Bao HF, Duke BJ, Liu BC, Ma HP, et al Cytochalasin E alters the cytoskeleton and decreases ENaC activity in Xenopus 2F3 cells. Am J Physiol Renal Physiol. 2014; 307(1): F86– 95. 10.1152/ajprenal.00251.2013 24829507 PMC4080154 · DOI
  24. [24]Cantiello HF, Stow JL, Prat AG, Ausiello DA. Actin filaments regulate epithelial Na+ channel activity. Am J Physiol. 1991; 261(5 Pt 1): C882– 8.. 1659214
  25. [25]Mazzochi C, Benos DJ, Smith PR. Interaction of epithelial ion channels with the actin-based cytoskeleton. Am J Physiol Renal Physiol. 2006; 291(6): F1113– 22. 10.1152/ajprenal.00195.2006. 16926444 · DOI
  26. [26]Mazzochi C, Bubien JK, Smith PR, Benos DJ. The carboxyl terminus of the alpha-subunit of the amiloride-sensitive epithelial sodium channel binds to F-actin. J Biol Chem. 2006; 281(10): 6528– 38. 10.1074/jbc.M509386200. 16356937 · DOI
  27. [27]Karpushev AV, Ilatovskaya DV, Pavlov TS, Negulyaev YA, Staruschenko A. Intact cytoskeleton is required for small G protein dependent activation of the epithelial Na+ channel. PLoS One. 2010; 5(1): e8827 10.1371/journal.pone.0008827 20098689 PMC2809106 · DOI
  28. [28]Alli AA, Bao HF, Alli AA, Aldrugh Y, Song JZ, Ma HP, et al Phosphatidylinositol phosphate-dependent regulation of Xenopus ENaC by MARCKS protein. Am J Physiol Renal Physiol. 2012; 303(6): F800– 11. 10.1152/ajprenal.00703.2011 22791334 PMC3468524 · DOI
  29. [29]Minaschek G, Groschel-Stewart U, Blum S, Bereiter-Hahn J. Microcompartmentation of glycolytic enzymes in cultured cells. Eur J Cell Biol. 1992; 58(2): 418– 28.. 1330571
  30. [30]Cueille N, Blanc CT, Riederer IM, Riederer BM. Microtubule-associated protein 1B binds glyceraldehyde-3-phosphate dehydrogenase. J Proteome Res. 2007; 6(7): 2640– 7. 10.1021/pr070081z. 17521179 · DOI
  31. [31]Patterson RL, van Rossum DB, Kaplin AI, Barrow RK, Snyder SH. Inositol 1,4,5-trisphosphate receptor/GAPDH complex augments Ca2+ release via locally derived NADH. Proc Natl Acad Sci U S A. 2005; 102(5): 1357– 9. 10.1073/pnas.0409657102 15677321 PMC547892 · DOI
  32. [32]Taruno A, Niisato N, Marunaka Y. Intracellular calcium plays a role as the second messenger of hypotonic stress in gene regulation of SGK1 and ENaC in renal epithelial A6 cells. Am J Physiol Renal Physiol. 2008; 294(1): F177– 86. 10.1152/ajprenal.00250.2007. 17959754 · DOI
  33. [33]Thai TL, Yu L, Galarza-Paez L, Wu MM, Lam HY, Bao HF, et al The Polarized Effect of Intracellular Calcium on the Renal Epithelial Sodium Channel Occurs as a Result of Subcellular Calcium Signaling Domains Maintained by Mitochondria. J Biol Chem. 2015; 290(48): 28805– 11. 10.1074/jbc.M115.668293 26451045 PMC4661396 · DOI
  34. [34]Jella KK, Rani S, O'Driscoll L, McClean B, Byrne HJ, Lyng FM. Exosomes are involved in mediating radiation induced bystander signaling in human keratinocyte cells. Radiat Res. 2014; 181(2): 138– 45. 10.1667/RR13337.1. 24502353 · DOI
  35. [35]Antonyak MA, Cerione RA. Emerging picture of the distinct traits and functions of microvesicles and exosomes. Proc Natl Acad Sci U S A. 2015; 112(12): 3589– 90. 10.1073/pnas.1502590112 25762069 PMC4378426 · DOI
  36. [36]Zeringer E, Barta T, Li M, Vlassov AV. Strategies for isolation of exosomes. Cold Spring Harb Protoc. 2015; 2015(4): 319– 23. 10.1101/pdb.top074476. 25834266 · DOI
  37. [37]Ma HP, Li L, Zhou ZH, Eaton DC, Warnock DG. ATP masks stretch activation of epithelial sodium channels in A6 distal nephron cells. Am J Physiol Renal Physiol. 2002; 282(3): F501– 5. 10.1152/ajprenal.00147.2001. 11832432 · DOI
  38. [38]Kumagai H, Sakai H. A porcine brain protein (35 K protein) which bundles microtubules and its identification as glyceraldehyde 3-phosphate dehydrogenase. J Biochem. 1983; 93(5): 1259– 69.. 6885722
  39. [39]Chen RW, Saunders PA, Wei H, Li Z, Seth P, Chuang DM. Involvement of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and p53 in neuronal apoptosis: evidence that GAPDH is upregulated by p53. J Neurosci. 1999; 19(21): 9654– 62.. 10531467 PMC6782921
  40. [40]Robbins AR, Ward RD, Oliver C. A mutation in glyceraldehyde 3-phosphate dehydrogenase alters endocytosis in CHO cells. J Cell Biol. 1995; 130(5): 1093– 104. 7657694 PMC2120546
  41. [41]Glaser PE, Gross RW. Rapid plasmenylethanolamine-selective fusion of membrane bilayers catalyzed by an isoform of glyceraldehyde-3-phosphate dehydrogenase: discrimination between glycolytic and fusogenic roles of individual isoforms. Biochemistry. 1995; 34(38): 12193– 203.. 7547960
  42. [42]Tisdale EJ. Glyceraldehyde-3-phosphate dehydrogenase is required for vesicular transport in the early secretory pathway. J Biol Chem. 2001; 276(4): 2480– 6. 10.1074/jbc.M007567200. 11035021 · DOI
  43. [44]Renigunta A, Mutig K, Rottermann K, Schlichthorl G, Preisig-Muller R, Daut J, et al The glycolytic enzymes glyceraldehyde 3-phosphate dehydrogenase and enolase interact with the renal epithelial K+ channel ROMK2 and regulate its function. Cell Physiol Biochem. 2011; 28(4): 663– 72. 10.1159/000335761. 22178878 · DOI

Cited by

Sorted by citations

Authors
Jella, K. K., Yu, L., Yue, Q., Friedman, D., Duke, B. J., & Alli, A. A.
Affiliations
  • Emory University (US)
  • Nanjing Agricultural University (CN)
  • University of Florida Health (US)
Corresponding author
Alli, Abdel A.
ORCID
Venue
PLOS ONE
Publisher
Public Library of Science (PLoS)
Published
2016-11-01
Volume
11
Issue
11
Pages
e0165763
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0165763
PMC
PMC5089749
OpenAlex
W2547591013
Cited by
81
References
1
Citations by year
2026: 2 · 2025: 3 · 2024: 8 · 2023: 8 · 2022: 17 · 2021: 11 · 2020: 10 · 2019: 8 · 2018: 4 · 2017: 9 · 2014: 1
FWCI
3.64
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
  • US National Aeronautics and Space Administration award (NNX15AD63G)
  • National Institutes of Health (K01 DK099617)
  • College of Medicine, University of Florida
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion Transport and Channel Regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
Keywords
exosomes, epithelial sodium channel, glyceraldehyde-3-phosphate dehydrogenase, proximal tubular epithelial cells, intercellular communication, exosome uptake, sodium reabsorption, renal epithelial cells
MeSH terms
Amino Acid Sequence, Animals, Cell Line, Female, Glyceraldehyde-3-Phosphate Dehydrogenases, Kidney Tubules, Proximal, Male, Protein Transport, Mice, Epithelial Sodium Channels, Exosomes

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