Rapid isolation of Tamm-Horsfall glycoprotein (uromodulin) from human urine

Serafini‐Cessi, Franca; Bellabarba, Giovanni; Malagolini, Nadia; Dall’Olio, Fabio

doi:10.1016/0022-1759(89)90241-x

Cited by 52 publications

(44 citation statements)

References 15 publications

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“…The pellet fraction was further purified by bringing the solution to 140 mM NaCl and 250 mM phosphate buffer, pH 7.5, which induces gel formation. The THP sample was then added onto a column containing diatomaceous earth, washed with PBS, and eluted by addition of H2O as described (24). The MAPK inhibitors SB203580, PD98059, and UO126 and the NF-κB inhibitor SN50 were purchased from Calbiochem.…”

Section: Methodsmentioning

confidence: 99%

“…The pellet fraction containing THP was fully active on C57BL/6 splenocytes for TNF-α and IL-6 production ( Figure 2D and data not shown). Moreover, the ultracentrifuged THP was further purified over a diatomaceous earth filter, which is known to purify THP to homogeneity (24). This THP preparation highly purified by 3 different methods was still able to induce TNF-α production ( Figure 2D).…”

Section: Thp Induces Phenotypic Maturation Of Human Monocyte-derived mentioning

confidence: 99%

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Tamm-Horsfall glycoprotein links innate immune cell activation with adaptive immunity via a Toll-like receptor-4–dependent mechanism

Säemann¹,

Weichhart²,

Zeyda³

et al. 2005

J. Clin. Invest.

200

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Tamm-Horsfall glycoprotein (THP) is expressed exclusively in the kidney and constitutes the most abundant protein in mammalian urine. A critical role for THP in antibacterial host defense and inflammatory disorders of the urogenital tract has been suggested. We demonstrate that THP activates myeloid DCs via Toll-like receptor-4 (TLR4) to acquire a fully mature DC phenotype. THP triggers typical TLR signaling, culminating in activation of NF-κB. Bone marrow-derived macrophages from TLR4-and MyD88-deficient mice were nonresponsive to THP in contrast to those from TLR2-and TLR9-deficient mice. In vivo THP-driven TNF-α production was evident in WT but not in Tlr4 -/-mice. Importantly, generation of THP-specific Abs consistently detectable in urinary tract inflammation was completely blunted in Tlr4 -/-mice. These data show that THP is a regulatory factor of innate and adaptive immunity and therefore could have significant impact on host immunity in the urinary tract.

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Section: Methodsmentioning

confidence: 99%

Section: Thp Induces Phenotypic Maturation Of Human Monocyte-derived mentioning

confidence: 99%

Tamm-Horsfall glycoprotein links innate immune cell activation with adaptive immunity via a Toll-like receptor-4–dependent mechanism

Säemann¹,

Weichhart²,

Zeyda³

et al. 2005

J. Clin. Invest.

200

View full text Add to dashboard Cite

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“…The ZP domain is highly species-conserved in all hitherto cloned THPs (21)(22)(23)(24). The DEF method used in this study to purify both hTHP and pTHP is based on the fact that when urine is filtered through a layer of diatomaceous earth, the THP polymers are selectively entrapped (35). THP polymers are solubilized in deionized water, so that by means of a two-step filtration at a concentration of NaCl of Ն0.15 M and by washing the diatomaceous earth with deionized water, THP is purified to homogeneity even from urine of proteinuric patients (50).…”

Section: Discussionmentioning

confidence: 99%

“…35 S-Labeled E. coli strains that express a low affinity variant of FimH (KB91) and a high affinity variant of FimH (KB96) were incubated with increasing amounts of immobilized BSA or monomeric pTHP and hTHP. Note that although there was little binding to BSA, both strains bound to THPs with KB96 bound with greater numbers and that both strains bound to pTHP approximately 3-fold higher than to hTHP.…”

Section: Figmentioning

confidence: 99%

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Variation of High Mannose Chains of Tamm-Horsfall Glycoprotein Confers Differential Binding to Type 1-fimbriated Escherichia coli

Cavallone

Malagolini

Monti

et al. 2004

Journal of Biological Chemistry

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Tamm-Horsfall glycoprotein (THP), the most abundant protein in mammalian urine, has been implicated in defending the urinary tract against infections by type 1-fimbriated Escherichia coli. Recent experimental evidence indicates that the defensive capability of THP relies on its single high mannose chain, which binds to E. coli FimH lectin and competes with mannosylated uroplakin receptors on the bladder surface. Here we describe several major differences, on both structural and functional levels, between human THP (hTHP) and pig THP (pTHP). pTHP contains a much higher proportion (47%) of Man 5 GlcNAc 2 than does hTHP (8%). FimHexpressing E. coli adhere to monomeric pTHP at an approximately 3-fold higher level than to monomeric hTHP. This suggests that the shorter high mannose chain (Man 5 GlcNAc 2 ) is a much better binder for FimH than the longer chains (Man 6 -7 GlcNAc 2 ) and that pTHP is a more potent urinary defense factor than hTHP. In addition, unlike hTHP whose polyantennary glycans are exclusively capped by sialic acid and sulfate groups, those of pTHP are also terminated by Gal␣1,3Gal epitope. This is consistent with the fact that the outer medulla of pig kidney expresses the ␣1,3-galactosyltransferase, which is completely absent in human kidney. Finally, pTHP is more resistant to leukocyte elastase hydrolysis than hTHP, thus explaining why pTHP is much less prone to urinary degradation than hTHP. These results demonstrate for the first time that the species variations of the glycomoiety of THP can lead to the differential binding of THP to type 1-fimbriated E. coli and that the differences in high mannose processing may reflect species-specific adaptation of urinary defenses against E. coli infections.

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Alterations of the sugar moiety of Tamm-Horsfall protein in children with malignancies of lymphoid cells

Olczak

Dereniowska

et al. 1999

Electrophoresis

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Rapid isolation of Tamm-Horsfall glycoprotein (uromodulin) from human urine

Cited by 52 publications

References 15 publications

Tamm-Horsfall glycoprotein links innate immune cell activation with adaptive immunity via a Toll-like receptor-4–dependent mechanism

Tamm-Horsfall glycoprotein links innate immune cell activation with adaptive immunity via a Toll-like receptor-4–dependent mechanism

Variation of High Mannose Chains of Tamm-Horsfall Glycoprotein Confers Differential Binding to Type 1-fimbriated Escherichia coli

Alterations of the sugar moiety of Tamm-Horsfall protein in children with malignancies of lymphoid cells

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