Serum antibodies to heat shock protein 70 in sensorineural hearing loss.
Serum antibodies to heat shock protein 70 in sensorineural hearing loss.
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DOI：
10.1001/archotol.1995.01890100075013
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发表时间：
1995-10
期刊：
影响因子：
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通讯作者：
D. Bloch;E. S. Martin;Steven D. Rauch;R. Moscicki;Kurt J. Bloch
中科院分区：
文献类型：
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作者：
D. Bloch;E. S. Martin;Steven D. Rauch;R. Moscicki;Kurt J. Bloch
目的鉴定进行性双耳感音神经性聋患者血清中抗体的68 kd靶点。设计采用凝胶过滤、离子交换层析和聚丙烯酰胺凝胶电泳从肾组织提取液中纯化目的蛋白，并转移到硝酸纤维素膜上。纯化的蛋白用胰蛋白酶消化，并通过高压液相色谱分离肽片段。结果经高效液相色谱分离得到一个分子量为2776的多肽组分。该肽内的22个氨基酸的序列与热休克蛋白70（HSP 70）的氨基酸424至445的序列相同。在Western印迹上，针对HSP 70的单克隆抗体（但能够识别组成型HSP 70 [HSC 70]和应激诱导型HSP 70）与纯化的68-kd蛋白反应。我们比较了6例特发性、进行性、双侧感音神经性耳聋患者的血清样本以及HSC 70单克隆抗体和HSP 70单克隆抗体与肾提取物的反应性。获得的模式表明，患者抗体优先针对HSP 70。结论进行性双耳感音神经性聋患者血清中抗体的靶点为HSP 70。
OBJECTIVE To identify the 68-kd target of antibody in serum samples from patients with idiopathic, progressive, bilateral sensorineural hearing loss. DESIGN To purify target protein from renal extracts using gel filtration, ion-exchange chromatography, and polyacrylamide gel electrophoresis and to transfer to nitrocellulose membranes. The purified protein was digested with trypsin, and peptide fragments were separated by high-pressure liquid chromatography. RESULTS One fraction obtained by high-performance liquid chromatography contained a peptide of 2776 molecular weight. The sequence of a stretch of 22 amino acids within this peptide was identical to that of amino acids 424 through 445 of heat shock protein 70 (HSP70). On Western blotting, monoclonal antibody directed against HSP70 (but capable of recognizing both constitutive HSP70 [HSC70] and stress-inducible HSP70) reacted with the purified 68-kd protein. We compared the reactivity of serum samples from six patients with idiopathic, progressive, bilateral sensorineural hearing loss, as well as monoclonal antibody to HSC70, and monoclonal antibody to HSP70 with renal extract. The pattern obtained suggested that patient antibodies are preferentially directed at HSP70. CONCLUSION The target of antibody in serum samples from patients with idiopathic, progressive, bilateral sensorineural hearing loss is HSP70.