A Multi-Site Study Confirms Abnormal Glycosylation in the Tamm-Horsfall Protein of Patients With Interstitial Cystitis
A Multi-Site Study Confirms Abnormal Glycosylation in the Tamm-Horsfall Protein of Patients With Interstitial Cystitis
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DOI：
10.1016/j.juro.2011.02.2699
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发表时间：
2011-07-01
影响因子：
6.6
通讯作者：
Argade, Sulabha
中科院分区：
文献类型：
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作者：
Parsons, C. Lowell;Proctor, Jeffrey;Argade, Sulabha
目的：我们证实了单一的网站观察减少唾液酸化和异常糖基化的Tamm-Horsfall蛋白质与间质性膀胱炎患者相比，对照subjects.Materials和方法：尿液样本从41个对照组和48例间质性膀胱炎患者从总共5个北美网站获得盲的方式参与者的状态。通过盐沉淀从尿液样品中分离Tamm-Horsfall蛋白。通过尺寸排阻色谱法测定蛋白质含量，并归一化为肌酸酐。通过1，2-二氨基-4，5-亚甲基二氧基苯（Sigma（R））高效液相色谱法和荧光检测器定量唾液酸。中性和氨基糖测定高pH阴离子交换色谱脉冲安培检测。用2-氨基苯甲酰胺标记N-聚糖，并使用高pH阴离子交换色谱法结合荧光检测进行分析。还通过基质辅助激光解吸/电离-飞行时间质谱法分析样品。全甲基化N-聚糖在质荷比为3,000 - 6,000范围内进行分析。结果：间质性膀胱炎患者Tamm-Horsfall蛋白的蛋白/肌酐比与对照组无差异（49.12 vs 46.4 mg/gm，p = 0.26）。与对照组相比，间质性膀胱炎患者的唾液酸含量（67与77 nmol/mg Tamm-Horsfall蛋白，p = 0.025）和总单糖含量（590.9与680.6 nmol/mg Tamm-Horsfall蛋白，p = 0.003）显著降低。结果得到2-氨基苯甲酰胺N-聚糖谱和质谱的支持，其显示主要四唾液酸化峰降低45（质荷比4，590）之间的Tamm-Horsfall蛋白质的差异。这些多位点数据证实了Tamm-1的异常糖基化。Horsfall蛋白发生在间质性膀胱炎患者中，可能在间质性膀胱炎病因中发挥作用。
Purpose: We confirm the single site observation of decreased sialylation and abnormal glycosylation of Tamm-Horsfall protein in patients with interstitial cystitis compared to control subjects.Materials and Methods: Urine samples from 41 controls and 48 patients with interstitial cystitis from a total of 5 North American sites were obtained in blinded fashion as to participant status. Tamm-Horsfall protein was isolated from urine samples by salt precipitation. Protein content was determined by size exclusion chromatography and normalized to creatinine. Sialic acid was quantified by 1,2-diamino-4,5-methylene dioxybenzene (Sigma (R)) high performance liquid chromatography with fluorescence detection. Neutral and amino sugars were determined by high pH anion exchange chromatography with pulsed amperometric detection. N-glycans were labeled with 2-aminobenzamide and profiled using high pH anion exchange chromatography with fluorescence detection. Samples were also analyzed by matrix assisted laser desorption/ionization-time of flight mass spectrometry. Permethylated N-glycans were analyzed in the mass-to-charge ratio range of 3,000 to 6,000.Results: There was no difference in the protein-to-creatinine ratio of Tamm-Horsfall protein from patients with interstitial cystitis vs controls (49.12 vs 46.4 mg/gm, p = 0.26). Sialic acid content (67 vs 77 nmol/mg Tamm-Horsfall protein, p = 0.025) and total monosaccharide content (590.9 vs 680.6 nmol/mg Tamm-Horsfall protein, p = 0.003) were significantly decreased in patients with interstitial cystitis vs controls. Results were supported by 2-aminobenzamide N-glycan profiling and mass spectrometry, which showed a 45% decrease in a major tetra-sialylated peak (mass-to-charge ratio 4,590) in Tamm-Horsfall protein from patients with interstitial cystitis compared to controls.Conclusions: These multisite data validate that abnormal glycosylation of Tamm-Horsfall protein occurs in patients with interstitial cystitis and may have a role in interstitial cystitis causation.