PHOSPHOLIPID HEAD GROUP RECOGNITION IN MEMBRANE BINDING PROTEIN ANNEXIN V

一种用于检测和鉴定的质谱仪方法 血浆蛋白转甲状腺素(TTR)的变异体 发展起来的。在Ttr中发现的单一氨基酸替代是 与家族性甲状腺激素转运蛋白淀粉样变性密切相关， 遗传性退行性疾病。ATTR的确诊性诊断 依赖于TTR变异体的检测和鉴定。这个 这里正在开发的方法是基于使用 免疫沉淀。变异体的检测是通过质量来实现的 电喷雾-质谱法测定完整蛋白质 (ESIN4S)。高效液相色谱/电喷雾电离质量 蛋白质胰酶消化的LC/ESIMS光谱分析 然后是随后的基质辅助激光解吸电离 飞行时间质谱仪和NLALDI POST源 相关回收的色谱组分的衰变(PSDMS) 含有变异肽的可用于鉴定未知的 变种。用ATTR的血清对该方法进行了成功的检验 具有已知变异(Va130-4Met和Va1122--&gt；Ile)的患者。一种新的TTR 用上述方法检测和鉴定了变异体Ser23-&gt；Asn 等电聚焦和限制性内切酶分析的方法 未能确定变种的性质。

A mass spectrometry approach for the detection and identification of variants of theplasma protein transthyretin (TTR) is being developed. The single amino acid substitutions found in TTR are closely associated with familial transthyretin amyloidosis (ATTR), a hereditary degenerative disease. A definitive diagnosis of ATTR relies on the detection and identification of TTRvariants. The approach being developed here is based on isolation of serum TTR using immunoprecipitation. The detection of the variant is achieved by mass measurement of the intact protein with electrospray mass spectrometry (ESIN4S). The liquid chromatography/electrospray ionization mass spectrometry (LC/ESIMS) analysis of the tryptic digest of the protein followed by subsequent matrix-assisted laser desorption ionization time-of-flight mass spectrometry (NLkLDI-TOFMS) and NLALDI post source decay (PSDMS) of the relevant recovered chromatographic fraction containing the variant peptide allow the identification of unknown variants. The method wassuccessfully tested using serum from ATTR patients with known variants (Va130-4Met and Va1122-->Ile). A new TTR variant, Ser23->Asn, was detected and identified using the above method where isoelectric focusing and restriction enzyme analysis failed to identify the nature of thevariant.