Abstract
Both Tamm‐Horsfall protein (THP) and collectin‐11 (CL‐11) are important molecules in acute kidney injury (AKI). In this study, we measured the change of glycosylation of THP in patients with AKI after surgery, using MALDI‐TOF MS and lectin array analysis. The amount of high‐mannose and core fucosylation in patients with AKI were higher than those in healthy controls. In vitro study showed that THP could bind to CL‐11 with affinity at 9.41 × 10−7 mol/L and inhibited activation of complement lectin pathway. The binding affinity decreased after removal of glycans on THP. Removal of fucose completely ablated the binding between the two proteins. While removal of high‐mannose or part of the N‐glycan decreased the binding ability to 30% or 60%. The results indicated that increase of fucose on THP played an important role via complement lectin pathway in AKI.
Highlights
Tamm-Horsfall protein (THP) is the most abundant protein in normal human urine, and carbohydrate forms 25% of the molecular mass of THP.[1]
2.3 | Binding of THP with CL-11, Mannose-binding lectin (MBL) and ficolins detected by enzyme-linked immunosorbent assay (ELISA)
We further explored whether the binding between THP and lectin pathway initiation molecules influenced the complement activity with a modified haemolysis assay
Summary
Tamm-Horsfall protein (THP) is the most abundant protein in normal human urine, and carbohydrate forms 25% of the molecular mass of THP.[1]. Collectin-11 activates lectin pathway via binding with fucose and mannose.[15] Removal of carbohydrate from THP reduced the ability of THP binding of C1q and eliminated the ability of THP to protect against complement activation.[16] abnormal glycosylation of THP was observed in patients with interstitial cystitis as well as in patients accepting renal transplantation,[17,18] and the disturbance of glycosylation influenced immunoreaction of THP, indicated that glycans on THP mediated the pathogenic process of urologic diseases.[18,19]. We first measured the binding between THP and CL-11 and explored whether the binding between the two molecules influenced the activation of complement system. 2.3 | Binding of THP with CL-11, Mannose-binding lectin (MBL) and ficolins detected by enzyme-linked immunosorbent assay (ELISA). The OD at 405 nm was measured using a microplate reader (iMark, BIO-RAD)
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