Identification of Human Uromodulin as the Tamm-Horsfall Urinary Glycoprotein

Pennica, Diane; Kohr, William J.; Kuang, Wun Jing; Glaister, Debbie; Aggarwal, Bharat B.; Chen, Ellson Y.; Goeddel, David V.

doi:10.1126/science.3453112

Cited by 259 publications

(160 citation statements)

References 33 publications

(8 reference statements)

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“…This protein has been purified to homogeneity by three different techniques of protein separation, namely preparative ultracentrifugation, native PAGE and size exclusion gel chromatography, with a subsequent SDS PAGE run in all three approaches. Based on sequence information derived from MALDI-TOF MS and recognition by a crossreacting polyclonal human THP antibody, this protein has been characterised to be closely related or identical to human THP and/or uromodulin [17].…”

Section: Discussionmentioning

confidence: 99%

Vitamin A excreted in the urine of canines is associated with a Tamm-Horsfall like protein

Schweigert¹,

Raila²,

Haebel

2002

Vet. Res.

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-Under physiological conditions canines transport vitamin A in blood plasma primarily as retinyl esters bound to lipoproteins and excrete substantial amounts of vitamin A as retinol and retinyl esters with urine. In the aqueous environment of urine, the hydrophobic vitamin A has to be associated with a protein. This vitamin A-protein complex was purified to homogeneity, prepared by preparative ultracentrifugation (density 1.21 g/mL), native polyacrylamide gel electrophoresis (PAGE) and size exclusion chromatography. The vitamin A-protein complex has a high molecular mass of > 5 000 kDa under native conditions. SDS PAGE under reduced conditions revealed a single band with a molecular mass of about 100 kDa for the protein moiety. Peptides obtained after limited proteolysis with trypsin from the 100 kDa protein were characterised by MALDI-TOF mass spectrometry and showed amino acid sequence homology to the human Tamm-Horsfall Protein (THP). This was further confirmed by a positive immunoreaction of the isolated protein with crossreacting human THP antibodies. The localisation of THP in dog kidneys was determined by using immunohistology. The reaction was strong along the entire thick ascending limb of the Henle loop and distal convoluted tubule. Our data point to the possibility that THP functions as a novel carrier for vitamin A in the urine of canines. l'environnement aqueux de l'urine, la vitamine A hydrophobe doit être associée à une protéine. Ce complexe protéine-vitamine A a été purifié et homogénéisé, préparé par ultracentrifugation (1,21 g/mL de densité), électrophorèse sur gel de polyacrylamide natif (PAGE) et chromatographie d'exclusion de taille. Le complexe protéine-vitamine A a une masse moléculaire élevée supérieure à 5 000 kDa dans les conditions natives. Le SDS PAGE effectué dans des conditions réduites a révélé une bande unique avec une masse moléculaire d'environ 100 kDa pour la partie protéique. Les peptides obtenus à partir de la protéine de 100 kDa, après protéolyse limitée avec de la trypsine, ont été caractérisés par spectrométrie de masse MALDI-TOF et ont montré une homologie de séquence en acides aminés avec la protéine Tamm-Horsfall (PTH) humaine. Ceci a été confirmé par une immuno-réaction positive entre la protéine isolée et les anticorps présentant une réaction croisée avec la PTH humaine. La localisation de la PTH dans le rein de chien a été déterminée par immunohistologie. La réaction a été forte tout le long de la section ascendante épaisse de l'anse de Henle, et du tubule contourné distal. Nos résultats suggèrent que la PTH fonctionne comme un nouveau transporteur pour la vitamine A dans l'urine des canidés.vitamine A / canidés / rein / urine / protéine Tamm-Horsfall

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Section: Discussionmentioning

confidence: 99%

Vitamin A excreted in the urine of canines is associated with a Tamm-Horsfall like protein

Schweigert¹,

Raila²,

Haebel

2002

Vet. Res.

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“…It is interesting that no mutation is predicted to result in premature termination of translation. The clustering of mutations in exon 4 is likely to be significant considering (1) a strong sequence conservation in evolution (38); (2) an approximately 53% sequence similarity with glycoprotein-2 (GP-2), a zymogen granule GPI-linked protein that, like uromodulin, is released from apical membranes to form large aggregates in solution (34,39); (3) the high number of cysteine residues in this part of the protein (24); and (4) the presence of three epidermal growth factor-like (EGF) repeats, able to interact with structurally related ligands (24,25,40) (Figure 1B). EGF domains represent one of the most commonly identified protein modules that mediate protein-protein interactions (40).…”

Section: Discussionmentioning

confidence: 99%

“…The UMOD gene encodes uromodulin, a 85-kd glycoprotein that is identical to the Tamm-Horsfall protein (24,25). Although uromodulin is the most abundant protein in the normal urine (33), its biologic role remains enigmatic.…”

Section: Discussionmentioning

confidence: 99%

“…(B) UMOD predicted protein structure: functional domains of UMOD are shown as shaded boxes, with their names. The N-terminal region (essentially exon 4) contains three epidermal growth factor (EGF)-like modules with typical Ca 2ϩ -binding consensus in two of them (cbEGF2 and cbEGF3) followed by a cysteine-rich sequence of 166 residues that are highly conserved in UMOD homologues from different species (24). The C-terminal region contains the zona pellucida (ZP), an approximately 260 -amino acid domain that is responsible for polymerization of this protein into filaments of similar supramolecular structure (26) and a phosphatidylinositol anchor (between codons Leu 601 and Ala 616 ) (25).…”

Section: Effect Of Umod Mutations On Urinary Excretion and Renal Exprmentioning

confidence: 99%

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A Cluster of Mutations in the UMOD Gene Causes Familial Juvenile Hyperuricemic Nephropathy with Abnormal Expression of Uromodulin

Dahan¹,

Devuyst²,

Smaers³

et al. 2003

Journal of the American Society of Nephrology

199

170

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Abstract. Familial juvenile hyperuricemic nephropathy (FJHN [MIM 162000]) is an autosomal-dominant disorder characterized by abnormal tubular handling of urate and late development of chronic interstitial nephritis leading to progressive renal failure. A locus for FJHN was previously identified on chromosome 16p12 close to the MCKD2 locus, which is responsible for a variety of autosomal-dominant medullary cystic kidney disease (MCKD2). UMOD, the gene encoding the Tamm-Horsfall/uromodulin protein, maps within the FJHN/MCKD2 critical region. Mutations in UMOD were recently reported in nine families with FJHN/ MCKD2 disease. A mutation in UMOD has been identified in 11 FJHN families (10 missense and one in-frame deletion)-10 of which are novel-clustering in the highly conserved exon 4. The consequences of UMOD mutations on uromodulin expression were investigated in urine samples and renal biopsies from nine patients in four families. There was a markedly increased expression of uromodulin in a cluster of tubule profiles, suggesting an accumulation of the protein in tubular cells. Consistent with this observation, urinary excretion of wild-type uromodulin was significantly decreased. The latter findings were not observed in patients with FJHN without UMOD mutations. In conclusion, this study points to a mutation clustering in exon 4 of UMOD as a major genetic defect in FJHN. Mutations in UMOD may critically affect the function of uromodulin, resulting in abnormal accumulation within tubular cells and reduced urinary excretion.Familial juvenile hyperuricemic nephropathy (FJHN) is an autosomal-dominant disorder characterized by hyperuricemia and decreased urinary excretion of urate, followed by the development of chronic interstitial nephritis most often leading to progressive renal failure (1,2). The link between early hyperuricemia and subsequent progression of renal disease remains unclear.Urate is the end product of purine metabolism in humans, who have lost the expression of the uricase gene during evolution (3). Urate is freely filtered by the glomerulus and essentially reabsorbed, because only 10% of the filtered load is present in the final urine (4). The transport mechanisms of urate are localized in the proximal tubule (PT), whereas no experimental evidence supports urate permeability in the more distal segments of the nephron (5). URAT1, the long-hypothesized apical urate-anion exchanger involved in the reabsorption of urate by PT cells, was recently identified (6). Inactivating mutations of URAT1 located on 11q13 are responsible

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“…In 1985, Muchmore and Decker isolated an 85-kDa glycoprotein with potent immunosuppressive properties from the urine of pregnant women and named it "uromodulin" to reflect its origin in the urine and its ability to inhibit antigen-induced T-cell proliferation and monocyte cytotoxicity in vitro [2]. This glycoprotein was subsequently confirmed to be THP by Pennica et al [3].…”

Section: Introductionmentioning

confidence: 99%