Effect of Cholesterol on the Organic Cation Transporter OCTN1 (SLC22A4)

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2020 Feb 12

PMID 32041338

Citations 5

Authors

Lorena Pochini

Gilda Pappacoda

Michele Galluccio

Francesco Pastore

Mariafrancesca Scalise

Cesare Indiveri

Affiliations

Soon will be listed here.

Abstract

The effect of cholesterol was investigated on the OCTN1 transport activity measured as [C]-tetraethylamonium or [H]-acetylcholine uptake in proteoliposomes reconstituted with native transporter extracted from HeLa cells or the human recombinant OCTN1 over-expressed in . Removal of cholesterol from the native transporter by MβCD before reconstitution led to impairment of transport activity. A similar activity impairment was observed after treatment of proteoliposomes harboring the recombinant (cholesterol-free) protein by MβCD, suggesting that the lipid mixture used for reconstitution contained some cholesterol. An enzymatic assay revealed the presence of 10 µg cholesterol/mg total lipids corresponding to 1% cholesterol in the phospholipid mixture used for the proteoliposome preparation. On the other way around, the activity of the recombinant OCTN1 was stimulated by adding the cholesterol analogue, CHS to the proteoliposome preparation. Optimal transport activity was detected in the presence of 83 µg CHS/ mg total lipids for both [C]-tetraethylamonium or [H]-acetylcholine uptake. Kinetic analysis of transport demonstrated that the stimulation of transport activity by CHS consisted in an increase of the Vmax of transport with no changes of the Km. Altogether, the data suggests a direct interaction of cholesterol with the protein. A further support to this interpretation was given by a docking analysis indicating the interaction of cholesterol with some protein sites corresponding to CARC-CRAC motifs. The observed direct interaction of cholesterol with OCTN1 points to a possible direct influence of cholesterol on tumor cells or on acetylcholine transport in neuronal and non-neuronal cells via OCTN1.

Citing Articles

Atomistic description of the OCTN1 recognition mechanism via in silico methods.

Ben Mariem O, Palazzolo L, Torre B, Wei Y, Bianchi D, Guerrini U PLoS One. 2024; 19(6):e0304512.

PMID: 38829838 PMC: 11146731. DOI: 10.1371/journal.pone.0304512.

Strategies for Successful Over-Expression of Human Membrane Transport Systems Using Bacterial Hosts: Future Perspectives.

Galluccio M, Console L, Pochini L, Scalise M, Giangregorio N, Indiveri C Int J Mol Sci. 2022; 23(7).

PMID: 35409183 PMC: 8998559. DOI: 10.3390/ijms23073823.

OCTN1: A Widely Studied but Still Enigmatic Organic Cation Transporter Linked to Human Pathology and Drug Interactions.

Pochini L, Galluccio M, Scalise M, Console L, Pappacoda G, Indiveri C Int J Mol Sci. 2022; 23(2).

PMID: 35055100 PMC: 8776198. DOI: 10.3390/ijms23020914.

Cholesterol stimulates the cellular uptake of L-carnitine by the carnitine/organic cation transporter novel 2 (OCTN2).

Zhang L, Gui T, Console L, Scalise M, Indiveri C, Hausler S J Biol Chem. 2020; 296:100204.

PMID: 33334877 PMC: 7948396. DOI: 10.1074/jbc.RA120.015175.

ATP modulates SLC7A5 (LAT1) synergistically with cholesterol.

Cosco J, Scalise M, Colas C, Galluccio M, Martini R, Rovella F Sci Rep. 2020; 10(1):16738.

PMID: 33028978 PMC: 7541457. DOI: 10.1038/s41598-020-73757-y.

References

Pochini L, Scalise M, Di Silvestre S, Belviso S, Pandolfi A, Arduini A . Acetylcholine and acetylcarnitine transport in peritoneum: Role of the SLC22A4 (OCTN1) transporter. Biochim Biophys Acta. 2016; 1858(4):653-60. DOI: 10.1016/j.bbamem.2015.12.026. View

Listowski M, Leluk J, Kraszewski S, Sikorski A . Cholesterol Interaction with the MAGUK Protein Family Member, MPP1, via CRAC and CRAC-Like Motifs: An In Silico Docking Analysis. PLoS One. 2015; 10(7):e0133141. PMC: 4505867. DOI: 10.1371/journal.pone.0133141. View

Drenberg C, Gibson A, Pounds S, Shi L, Rhinehart D, Li L . OCTN1 Is a High-Affinity Carrier of Nucleoside Analogues. Cancer Res. 2017; 77(8):2102-2111. PMC: 5419029. DOI: 10.1158/0008-5472.CAN-16-2548. View

Scalise M, Galluccio M, Pochini L, Cosco J, Trotta M, Rebsamen M . Insights into the transport side of the human SLC38A9 transceptor. Biochim Biophys Acta Biomembr. 2019; 1861(9):1558-1567. DOI: 10.1016/j.bbamem.2019.07.006. View

Kulig W, Jurkiewicz P, Olzynska A, Tynkkynen J, Javanainen M, Manna M . Experimental determination and computational interpretation of biophysical properties of lipid bilayers enriched by cholesteryl hemisuccinate. Biochim Biophys Acta. 2014; 1848(2):422-32. DOI: 10.1016/j.bbamem.2014.10.032. View

Tamai I, Nakanishi T, Kobayashi D, China K, Kosugi Y, Nezu J . Involvement of OCTN1 (SLC22A4) in pH-dependent transport of organic cations. Mol Pharm. 2005; 1(1):57-66. DOI: 10.1021/mp0340082. View

Tamai I, Ohashi R, Nezu J, Sai Y, Kobayashi D, Oku A . Molecular and functional characterization of organic cation/carnitine transporter family in mice. J Biol Chem. 2000; 275(51):40064-72. DOI: 10.1074/jbc.M005340200. View

Castellano B, Thelen A, Moldavski O, Feltes M, van der Welle R, Mydock-McGrane L . Lysosomal cholesterol activates mTORC1 via an SLC38A9-Niemann-Pick C1 signaling complex. Science. 2017; 355(6331):1306-1311. PMC: 5823611. DOI: 10.1126/science.aag1417. View

Kato Y, Kubo Y, Iwata D, Kato S, Sudo T, Sugiura T . Gene knockout and metabolome analysis of carnitine/organic cation transporter OCTN1. Pharm Res. 2010; 27(5):832-40. DOI: 10.1007/s11095-010-0076-z. View

10.

Ishimoto T, Nakamichi N, Nishijima H, Masuo Y, Kato Y . Carnitine/Organic Cation Transporter OCTN1 Negatively Regulates Activation in Murine Cultured Microglial Cells. Neurochem Res. 2017; 43(1):116-128. DOI: 10.1007/s11064-017-2350-5. View

11.

Tamai I . Pharmacological and pathophysiological roles of carnitine/organic cation transporters (OCTNs: SLC22A4, SLC22A5 and Slc22a21). Biopharm Drug Dispos. 2012; 34(1):29-44. DOI: 10.1002/bdd.1816. View

12.

Grundemann D, Harlfinger S, Golz S, Geerts A, Lazar A, Berkels R . Discovery of the ergothioneine transporter. Proc Natl Acad Sci U S A. 2005; 102(14):5256-61. PMC: 555966. DOI: 10.1073/pnas.0408624102. View

13.

Masuo Y, Ohba Y, Yamada K, Al-Shammari A, Seba N, Nakamichi N . Combination Metabolomics Approach for Identifying Endogenous Substrates of Carnitine/Organic Cation Transporter OCTN1. Pharm Res. 2018; 35(11):224. DOI: 10.1007/s11095-018-2507-1. View

14.

Yang X, Ma Z, Zhou S, Weng Y, Lei H, Zeng S . Multiple Drug Transporters Are Involved in Renal Secretion of Entecavir. Antimicrob Agents Chemother. 2016; 60(10):6260-70. PMC: 5038284. DOI: 10.1128/AAC.00986-16. View

15.

Mihaljevic I, Popovic M, Zaja R, Smital T . Phylogenetic, syntenic, and tissue expression analysis of slc22 genes in zebrafish (Danio rerio). BMC Genomics. 2016; 17(1):626. PMC: 4982206. DOI: 10.1186/s12864-016-2981-y. View

16.

Shinozaki Y, Furuichi K, Toyama T, Kitajima S, Hara A, Iwata Y . Impairment of the carnitine/organic cation transporter 1-ergothioneine axis is mediated by intestinal transporter dysfunction in chronic kidney disease. Kidney Int. 2017; 92(6):1356-1369. DOI: 10.1016/j.kint.2017.04.032. View

17.

Pochini L, Scalise M, Galluccio M, Amelio L, Indiveri C . Reconstitution in liposomes of the functionally active human OCTN1 (SLC22A4) transporter overexpressed in Escherichia coli. Biochem J. 2011; 439(2):227-33. DOI: 10.1042/BJ20110544. View

18.

Pettersen E, Goddard T, Huang C, Couch G, Greenblatt D, Meng E . UCSF Chimera--a visualization system for exploratory research and analysis. J Comput Chem. 2004; 25(13):1605-12. DOI: 10.1002/jcc.20084. View

19.

Scanlon S, Williams D, Schloss P . Membrane cholesterol modulates serotonin transporter activity. Biochemistry. 2001; 40(35):10507-13. DOI: 10.1021/bi010730z. View

20.

Scalise M, Pochini L, Cosco J, Aloe E, Mazza T, Console L . Interaction of Cholesterol With the Human SLC1A5 (ASCT2): Insights Into Structure/Function Relationships. Front Mol Biosci. 2019; 6:110. PMC: 6819821. DOI: 10.3389/fmolb.2019.00110. View