人类APPL2蛋白PTB结构域的表达纯化及其功能的研究

APPL蛋白,包括APPL1和APPL2,是细胞膜与细胞核之间重要的信息传递者,同时也为细胞增殖所必需。在大肠杆菌表达体系中,表达并纯化了人类APPL2蛋白PTB结构域,然后以此为靶标,利用噬菌体展示技术筛选相互作用肽段。经过3轮筛选,随机挑取48个噬菌体单克隆,进行ELISA分析。7肽序列ERLPFFY和YLTSPKH被证明能与PTB结构域特异地结合。对P3GST的野生型和将每个氨基酸突变为丙氨酸的突变型的ELISA检测,显示每个氨基酸均是结合所必需的。这些对有关APPL2的PTB结构域功能的进一步研究有参考意义。     

2型糖尿病患者APPL1、AFABP与胰岛素抵抗指数的相关性研究

目的:分析2型糖尿病(T2DM)患者磷酸酪氨酸衔接蛋白(APPL1)、脂肪细胞型脂肪酸结合蛋白(AFABP)与稳态模型评估胰岛素抵抗指数(HOMA-IR)的相关性。方法:选择2015年6月~2016年5月至我院就诊T2DM患者100例作为患病组,选取同期在我院健康体检者100例作为健康组,对研究对象进行指标如空腹血糖(FPG)、空腹血清胰岛素(FINS)、糖化血红蛋白(Hb A1c)、总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)、APPL1、AFABP等检测,并根据公式计算HOMA-IR、及体重指数(BMI),分析APPL1、AFABP与各指标相关性。结果:患病组与健康组TC、HDL、LDL水平无明显差异(P0.05),患病组BMI、FPG、FINS、Hb A1c、TG、HOMA-IR、APPL1、AFABP与健康组比较明显较高(P0.05);APPL1与BMI、FINS、Hb A1c、HOMA-IR呈负相关性(P0.05),与FPG呈正相关性;AFABP与BMI、FPG、FINS、Hb A1c、HOMA-IR呈正相关性(P0.05)。结论:T2DM患者APPL1、AFABP较高,APPL1、AFABP与HOMA-IR呈直线相关性,表明APPL1、AFABP与T2DM患者胰岛素抵抗密切相关,该研究为APPL1、AFABP可以作为T2DM治疗的新靶点提供了理论依据。     

The PDZ Protein GIPC Regulates Trafficking of the LPA1 Receptor from APPL Signaling Endosomes and Attenuates the Cell’s Response to LPA

Lysophosphatidic acid (LPA) mediates diverse cellular responses through the activation of at least six LPA receptors – LPA_1–6, but the interacting proteins and signaling pathways that mediate the specificity of these receptors are largely unknown. We noticed that LPA₁ contains a PDZ binding motif (SVV) identical to that present in two other proteins that interact with the PDZ protein GIPC. GIPC is involved in endocytic trafficking of several receptors including TrkA, VEGFR2, lutropin and dopamine D2 receptors. Here we show that GIPC binds directly to the PDZ binding motif of LPA₁ but not that of other LPA receptors. LPA₁ colocalizes and coimmunoprecipitates with GIPC and its binding partner APPL, an activator of Akt signaling found on APPL signaling endosomes. GIPC depletion by siRNA disturbed trafficking of LPA₁ to EEA1 early endosomes and promoted LPA₁ mediated Akt signaling, cell proliferation, and cell motility. We propose that GIPC binds LPA₁ and promotes its trafficking from APPL-containing signaling endosomes to EEA1 early endosomes and thus attenuates LPA-mediated Akt signaling from APPL endosomes.