摘要
目的:探讨转甲状腺素(TTR)蛋白代谢修饰与淀粉样变形成的分子机制。方法:本研究首先利用ProteomeLabPF-2D系统依据蛋白质等电点分选各个p H值范围蛋白质,然后利用抗人TTR抗体,点免疫印迹实现鉴定TTR蛋白存在于被收集的范围。利用硫黄素T(ThT)试剂,测定各个收集部分溶液中淀粉样变的含量,进一步利用飞行质谱的表面增强激光解吸/电离时间(SELDI-TOF-MS)分析各个部分TTR蛋白的修饰类型。结果:系统性老年性淀粉样变(SSA)患者和健康志愿者血清pH范围分别为8.1～8.0,4.0～4.6及6.8～5.8范围时出现高丰度蛋白。并且SSA患者p H6.8～5.8范围淀粉样变含量明显高于对照组(P<0.05),其中SSA组与健康组相比,血TTR蛋白N端氨基酸修饰基团包括磷酰基(Phosphoryl)、羟基(Hydroxyl)、葡萄糖胺(O-GlcNac)基团修饰阴性。结论:SSA患者血TTR蛋白N端氨基酸修饰基团Phosphoryl、Hydroxyl、O-GlcNac的减少可能与淀粉样蛋白的形成有关。
Objective: To explore TTR event protein metabolism and the molecular mechanism of amyloid formation. Methods: We first used ProteomeLabPF - 2D system to separate protein on the basis of protein isoelectric point(IP) in each pH range and then western blot was applied to identify the TTR range. We used sulfur flavin T (ThT) reagent quantitative to sort out the collection part of the content of amyloidosis, further flight mass spectrometry of surface-enhanced laser desorption/ionization- time (SELDI TOF MS) to analyze each part of TTR event protein modification type. Results: It was found that SSA patients and healthy volunteers whose serum pH were 8.0- 8.6, pH 4.0-4.6 and pH 5.8-6.8 had a high abundance of protein. Moreover, during pH 6.8 to 5.8, the amyloidosis content of SSA patient group had significantly higher pH than the control group. The TTR N-terminal amino acid modified groups: Phosphoryl, Hydroxyl, O - GlcNac were negative. Conclusion: The reduction of TTR event N-terminal amino acid modified protein was found in SSA patients and Phosphoryl (Phosphoryl), Hydroxyl (Hydroxyl), glucosamine (O - GlcNac) may be associated with the formation of amyloid protein.
作者
赵莉逦
刘晓丹
PUREVDORJ Narangerel
王然
李蕾
孙续国
ZHAO Li-li;LIU Xiao-dan;PUREVDORJ Narangerel;WANG Ran;LI Lei;SUN Xu-guo(Department of Clinical Laboratory and Blood, School of Medical Laboratory, Tianjin Medical University, Tianjin 300203, China)
出处
《天津医科大学学报》
2019年第1期10-14,共5页
Journal of Tianjin Medical University
基金
国家自然科学基金资助项目(30973157)
天津市科学技术基金国家资助(15JCYBJC268000)
