Expression, purification and fluorine-18 radiolabeling of recombinant S100 proteins -: potential probes for molecular imaging of receptor for advanced glycation endproducts (RAGE) in vivo
Expression, purification and fluorine-18 radiolabeling of recombinant S100 proteins -: potential probes for molecular imaging of receptor for advanced glycation endproducts (RAGE) in vivo
复制标题
DOI：
10.1016/j.pep.2007.10.009
复制
发表时间：
2008-02-01
影响因子：
1.6
通讯作者：
Pletzsch, Jens
中科院分区：
文献类型：
--
作者：
Hoppmann, Susan;Haase, Cathleen;Pletzsch, Jens
循环S100蛋白（Ca2+调节蛋白的多基因家族）与晚期糖基化终产物受体（RAGE）在心血管疾病、炎症过程和肿瘤发生中的相互作用的病理生理作用的数据。Vivo是稀缺的。原因之一是缺乏合适的放射性示踪剂方法。我们报告了一种新的方法，使用重组人S100A1， S100B和S100A12作为这种相互作用的分子成像的潜在探针。因此，将人S100蛋白克隆为GST融合蛋白，在细菌表达载体pGEX-6P-1中表达，并在大肠杆菌菌株BL21中表达。纯化的重组人S100蛋白用正电子发射器氟-18 （F-18）与n -琥珀酰酰-4-[F-18]氟苯甲酸盐（[F-18]SFB）偶联进行放射性标记。放射性标记重组S100蛋白（F-18-S100）用于大鼠生物分布实验和小动物正电子发射断层扫描（PET）研究。F-18-S100蛋白在体内的组织特异性分布与RAGE的解剖定位（如肺和血管系统）密切相关。这些发现表明，在大鼠体内循环的S100A1、S100B和S100A12蛋白是RAGE的配体。该方法允许使用小动物PET，并提供新的探针来描述正常和病理生理条件下RAGE在啮齿动物疾病模型中的功能表达。(C) 2007爱思唯尔公司版权所有。
Data concerning the pathophysiological role of the interaction of circulating S100 proteins, a multigenic family of Ca2+-modulated proteins, with the receptor for advanced glycation endproducts (RAGE) in cardiovascular diseases, inflammatory processes, and tumorigenesis it? vivo are scarce. One reason is the shortage of suitable radiotracer methods. We report a novel methodology using recombinant human S100A1, S100B, and S100A12 as potential probes for molecular imaging of this interaction. Therefore, human S100 proteins were cloned as GST fusion proteins in the bacterial expression vector pGEX-6P-1 and expressed in E. coli strain BL21. Purified recombinant human S100 proteins were radiolabeled with the positron emitter fluorine-18 (F-18) by conjugation with N-succinimidyl-4-[F-18]fluorobenzoate ([F-18]SFB). The radiolabeled recombinant S100 proteins (F-18-S100) were used in biodistribution experiments and small animal positron emission tomography (PET) studies in rats. The tissue-specific distribution of F-18-S100 proteins in vivo correlated well with the anatomical localization of RAGE, e.g., in lungs and in the vascular system. These findings indicate circulating S100A1, S100B, and S100A12 proteins to be ligands for RAGE in rats in vivo. The approach allows the use of small animal PET and provides novel probes to delineate functional expression of RAGE under normal and pathophysiological conditions in rodent models of disease. (C) 2007 Elsevier Inc. All rights reserved.