title: Lipocalin-2 (24p3/neutrophil gelatinase-associated lipocalin (NGAL)) receptor is expressed in distal nephron and mediates protein endocytosis.
abstract: In the kidney, bulk reabsorption of filtered proteins occurs in the proximal tubule via receptor-mediated endocytosis (RME) through the multiligand receptor complex megalin-cubilin. Other mechanisms and nephron sites for RME of proteins are unclear. Recently, the secreted protein 24p3 (lipocalin-2, neutrophil gelatinase-associated lipocalin (NGAL)), which is expressed in the distal nephron, has been identified as a sensitive biomarker of kidney damage. A high-affinity receptor for 24p3 (24p3R) that is involved in endocytotic iron delivery has also been cloned. We investigated the localization of 24p3R in rodent kidney and its role in RME of protein-metal complexes and albumin. Immunostaining of kidney tissue showed expression of 24p3R in apical membranes of distal tubules and collecting ducts, but not of proximal tubule. The differential expression of 24p3R in these nephron segments was confirmed in the respective cell lines. CHO cells transiently transfected with 24p3R or distal tubule cells internalized submicromolar concentrations of fluorescence-coupled proteins transferrin, albumin, or metallothionein (MT) as well as the toxic cadmium-MT (Cd2+(7)-MT) complex, which caused cell death. Uptake of MT or transferrin and Cd2+(7)-MT toxicity were prevented by picomolar concentrations of 24p3. An EC50 of 123±50 nM was determined for binding of MT to 24p3R by microscale thermophoresis. Hence, 24p3R binds proteins filtered by the kidney with high affinity and may contribute to RME of proteins, including 24p3, and to Cd2+(7)-MT toxicity in distal nephron segments.
authors.affiliation: Institute of Physiology and Pathophysiology, Centre for Biomedical Training and Research (ZBAF), University of Witten/Herdecke, 58453 Witten, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Langelueddecke
authors.affiliation: Institute of Anatomy and Cell Biology II, Albert-Ludwigs-Universität Freiburg, 79104 Freiburg, Germany.
authors.forename: Eleni
authors.initials: E
authors.lastname: Roussa
authors.affiliation: Department of Biomedicine, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Fenton
authors.affiliation: Institute of Physiology and Pathophysiology, Centre for Biomedical Training and Research (ZBAF), University of Witten/Herdecke, 58453 Witten, Germany.
authors.forename: Natascha A
authors.initials: NA
authors.lastname: Wolff
authors.affiliation: Institute of Physiology and Pathophysiology, Centre for Biomedical Training and Research (ZBAF), University of Witten/Herdecke, 58453 Witten, Germany.
authors.forename: Wing-Kee
authors.initials: WK
authors.lastname: Lee
authors.affiliation: Institute of Physiology and Pathophysiology, Centre for Biomedical Training and Research (ZBAF), University of Witten/Herdecke, 58453 Witten, Germany. Electronic address: frank.thevenod@uni-wh.de.
authors.forename: Frank
authors.initials: F
authors.lastname: Thévenod
mesh_terms: D000209:Acute-Phase Proteins / Q000378:metabolism*; D000418:Albumins / Q000378:metabolism; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004705:Endocytosis / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D007679:Kidney Medulla / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D007686:Kidney Tubules, Distal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D000071068:Lipocalin-2; D054834:Lipocalins / Q000378:metabolism*; D008297:Male; D008668:Metallothionein / Q000633:toxicity; D015513:Oncogene Proteins / Q000378:metabolism*; D011806:Quinolinium Compounds / Q000378:metabolism; D051381:Rats; D014168:Transferrin / Q000378:metabolism
chemical_list: D000209:Acute-Phase Proteins; D000418:Albumins; C481052:Alexa fluor 546; D000071068:Lipocalin-2; D054834:Lipocalins; D015513:Oncogene Proteins; D011806:Quinolinium Compounds; D014168:Transferrin; C004203:cadmium-binding protein; C071332:Lcn2 protein, mouse; D008668:Metallothionein
references.citation: J Clin Invest. 2005 Mar;115(3):610-21
references.pmid: 15711640
references.citation: J Clin Invest. 2010 Nov;120(11):4065-76
references.pmid: 20921623
references.citation: Anal Biochem. 1976 May 7;72:248-54
references.pmid: 942051
references.citation: J Cell Biol. 1987 Jul;105(1):207-14
references.pmid: 3611186
references.citation: Mol Cell. 2002 Nov;10(5):1033-43
references.pmid: 12453412
references.citation: Nat Rev Mol Cell Biol. 2002 Apr;3(4):256-66
references.pmid: 11994745
references.citation: Am J Physiol Renal Physiol. 2004 Sep;287(3):F393-403
references.pmid: 15126248
references.citation: Am J Physiol Regul Integr Comp Physiol. 2007 Dec;293(6):R2400-11
references.pmid: 17855492
references.citation: J Cell Mol Med. 2011 Feb;15(2):209-19
references.pmid: 20015204
references.citation: Am J Physiol. 1970 Aug;219(2):358-63
references.pmid: 5448064
references.citation: Angew Chem Int Ed Engl. 2010 Mar 15;49(12):2238-41
references.pmid: 20186894
references.citation: FEBS Lett. 2005 Jan 31;579(3):773-7
references.pmid: 15670845
references.citation: Kidney Int. 1979 Sep;16(3):247-50
references.pmid: 529675
references.citation: Kidney Int. 2006 Feb;69(3):440-9
references.pmid: 16514429
references.citation: Kidney Int. 1982 Feb;21(2):354-61
references.pmid: 7069998
references.citation: Am J Physiol Renal Physiol. 2009 Apr;296(4):F902-11
references.pmid: 19193726
references.citation: Am J Physiol. 1988 Oct;255(4 Pt 1):G429-40
references.pmid: 3263053
references.citation: Kidney Int. 2009 Feb;75(3):295-303
references.pmid: 18843259
references.citation: Toxicol Sci. 2011 Jun;121(2):343-56
references.pmid: 21436125
references.citation: Science. 2001 Aug 3;293(5531):829-34
references.pmid: 11486081
references.citation: J Histochem Cytochem. 1964 Jun;12:470-80
references.pmid: 14200765
references.citation: J Biol Chem. 1978 Mar 25;253(6):1930-7
references.pmid: 204636
references.citation: Am J Physiol Renal Physiol. 2007 Sep;293(3):F705-12
references.pmid: 17596526
references.citation: Am J Physiol Renal Physiol. 2001 Dec;281(6):F1028-35
references.pmid: 11704553
references.citation: Kidney Int. 2001 Nov;60(5):1885-92
references.pmid: 11703607
references.citation: J Histochem Cytochem. 1972 Feb;20(2):89-96
references.pmid: 4110528
references.citation: J Biol Chem. 2011 Jun 10;286(23):20606-14
references.pmid: 21507940
references.citation: Cell. 2010 Jun 11;141(6):1006-17
references.pmid: 20550936
references.citation: J Am Soc Nephrol. 2003 Oct;14(10):2534-43
references.pmid: 14514731
references.citation: Am J Physiol Renal Physiol. 2006 Jun;290(6):F1525-33
references.pmid: 16449358
references.citation: Biometals. 2010 Oct;23(5):897-926
references.pmid: 20549307
references.citation: J Biol Chem. 2000 Jul 21;275(29):22220-8
references.pmid: 10751401
references.citation: Cell. 2005 Dec 29;123(7):1293-305
references.pmid: 16377569
references.citation: Prog Food Nutr Sci. 1987;11(1):1-37
references.pmid: 3303133
references.citation: Toxicol Lett. 1989 Mar;46(1-3):13-29
references.pmid: 2650020
references.citation: Pflugers Arch. 2009 Oct;458(6):1039-48
references.pmid: 19499243
references.citation: Toxicology. 1989 Apr;55(1-2):117-29
references.pmid: 2652377
references.citation: J Clin Invest. 1970 Jan;49(1):1-4
references.pmid: 5409805
references.citation: Am J Physiol Cell Physiol. 2003 Dec;285(6):C1367-76
references.pmid: 12890648
references.citation: Proc Natl Acad Sci U S A. 2006 Feb 7;103(6):1834-9
references.pmid: 16446425
references.citation: Am J Physiol Renal Physiol. 2001 Dec;281(6):F1021-7
references.pmid: 11704552
references.citation: Biochem J. 1982 Oct 15;208(1):19-26
references.pmid: 6297460
references.citation: Kidney Int. 2008 May;73(9):994-1007
references.pmid: 18272962
references.citation: Proc Natl Acad Sci U S A. 2006 Dec 26;103(52):19678-82
references.pmid: 17164337
references.citation: Mol Cell. 2002 Nov;10(5):1045-56
references.pmid: 12453413
references.citation: Nephron Physiol. 2003;93(4):p87-93
references.pmid: 12759569
references.citation: Kidney Int. 1979 Sep;16(3):394-403
references.pmid: 529685
references.citation: J Clin Invest. 1992 Aug;90(2):429-38
references.pmid: 1322939
references.citation: Nat Med. 2011 Feb;17(2):216-22
references.pmid: 21240264
references.citation: Biochim Biophys Acta. 1971 Jan 26;230(1):96-104
references.pmid: 5543335
references.citation: Toxicol Lett. 2011 Jun 24;203(3):210-8
references.pmid: 21421027
references.citation: Nature. 2004 Dec 16;432(7019):917-21
references.pmid: 15531878
references.citation: J Pharmacol Exp Ther. 2006 Aug;318(2):782-91
references.pmid: 16690719
references.citation: Biometals. 2010 Oct;23(5):783-92
references.pmid: 20354761
references.citation: J Am Soc Nephrol. 2007 Feb;18(2):407-13
references.pmid: 17229907
references.citation: Pediatr Nephrol. 2010 May;25(5):813-22
references.pmid: 20049615
references.citation: Environ Health Perspect. 1984 Mar;54:249-66
references.pmid: 6734560
references.citation: Am J Physiol. 1992 Oct;263(4 Pt 2):F601-6
references.pmid: 1415732
references.citation: Nat Commun. 2010 Oct 19;1:100
references.pmid: 20981028
references.citation: Toxicol Appl Pharmacol. 2009 Aug 1;238(3):221-39
references.pmid: 19371614
references.citation: Nat Chem Biol. 2010 Aug;6(8):602-9
references.pmid: 20581821