An antiproliferative factor from interstitial cystitis patients is a frizzled 8 protein-related sialoglycopeptide
An antiproliferative factor from interstitial cystitis patients is a frizzled 8 protein-related sialoglycopeptide
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DOI：
10.1073/pnas.0404509101
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发表时间：
2004-08-10
影响因子：
11.1
通讯作者：
Michejda, CJ
中科院分区：
文献类型：
--
作者：
Keay, SK;Szekely, Z;Michejda, CJ
美国约有100万人患有间质性膀胱炎，这是一种慢性疼痛性膀胱疾病，其特征在于膀胱上皮衬里变薄或溃疡;其病因不明。我们已经确定了一种糖基化卷曲相关肽抑制细胞增殖，是分泌特异性膀胱上皮细胞从这种疾病的患者。这种抗增殖因子（APF）通过调节细胞粘附蛋白和生长因子的产生来深刻地抑制膀胱细胞增殖。APF的结构通过使用离子阱质谱（MS）、酶消化、凝集素亲和色谱和全合成来推断，并且通过在微毛细管反相液相色谱（muRPLC）/MS上共洗脱天然和合成的APF衍生物来证实。热稳定的唾液酸糖肽，其肽链与frizzled 8的推定的第六跨膜结构域具有100%同源性。合成的APF与天然的APF在正常膀胱上皮细胞和T24膀胱癌细胞中具有相同的生物学活性。北方印迹分析表明，含有卷曲8片段序列的探针与从间质性膀胱炎患者细胞中提取的mRNA结合，但不与对照结合。因此，APF是一种卷曲相关肽生长抑制剂，仅含有卷曲蛋白的跨膜片段，是间质性膀胱炎的潜在生物标志物。
Approximately 1 million people in the United States suffer from interstitial cystitis, a chronic painful urinary bladder disorder characterized by thinning or ulceration of the bladder epithelial lining; its etiology is unknown. We have identified a glycosylated frizzled-related peptide inhibitor of cell proliferation that is secreted specifically by bladder epithelial cells from patients with this disorder. This antiproliferative factor (APF) profoundly inhibits bladder cell proliferation by means of regulation of cell adhesion protein and growth factor production. The structure of APF was deduced by using ion trap mass spectrometry (MS), enzymatic digestion, lectin affinity chromatography, and total synthesis, and confirmed by coelution of native and synthetic APF derivatives on microcapillary reversed-phase liquid chromatography (muRPLC)/MS. APF was determined to be an acidic, heat-stable sialoglycopeptide whose peptide chain has 100% homology to the putative sixth transmembrane domain of frizzled 8. Both synthetic and native APF had identical biological activity in normal bladder epithelial cells and T24 bladder cancer cells. Northern blot analysis indicated binding of a probe containing the sequence for the frizzled 8 segment with mRNA extracted from cells of patients with interstitial cystitis but not controls. APF is therefore a frizzled-related peptide growth inhibitor shown to contain exclusively a transmembrane segment of a frizzled protein and is a potential biomarker for interstitial cystitis.