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首页> 外文期刊>Biochemistry >Strain-based Sequence Variations and Structure Analysis of Murine Prostate Specific Spermine Binding Protein Using Mass Spectrometry
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Strain-based Sequence Variations and Structure Analysis of Murine Prostate Specific Spermine Binding Protein Using Mass Spectrometry

机译:质谱基于小鼠的应变序列变异和小鼠前列腺特异性精胺结合蛋白的结构分析

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摘要

Mouse spermine binding protein (SBP) has been characterized using mass spectrometry, including its localization within the prostate, sequence verification, and its posttranslational modifications. MALDI (matrix-assisted laser desorption/ionization) mass spectrometry was employed for localization of proteins expressed by different lobes of the mouse prostate obtained after tissue blotting on a polyethylene membrane. The mass spectra showed complex protein profiles that were different for each lobe of the prostate. The prostate-specific spermine binding protein (SBP), primarily identified by its in-source decay fragment ion signals, was found predominantly expressed by the ventral lobe of the prostate. The MALDI in-source decay measurements combined with nanoESI (nanoelectrospay ionization) MS/MS measurements obtained after specific proteolysis of SBP, allowed the exact positioning of a single N-Iinked carbohydrate group, and the identification of a pyroglutamate residue at the sequence N-terminus. The N-Iinked carbohydrate component was further investigated and the general pattern of the N-Iinked carbohydrate identified. The presence of a disulfide bridge between cysteine78 and cysteinel24 was also established. The full sequence characterization of SBP showed several strain-based sequence differences when compared to the published gene sequence.
机译:小鼠精胺结合蛋白(SBP)已通过质谱法进行了表征,包括其在前列腺中的定位,序列验证及其翻译后修饰。 MALDI(基质辅助激光解吸/电离)质谱法用于定位由组织印迹在聚乙烯膜上后获得的小鼠前列腺的不同叶表达的蛋白质。质谱图显示了复杂的蛋白质谱,该蛋白谱对于前列腺的每个叶都是不同的。前列腺特异性精胺结合蛋白(SBP)主要由其源内衰变片段离子信号确定,主要由前列腺的腹叶表达。在对SBP进行特定蛋白水解后获得的MALDI源衰减测量值与nanoESI(纳米电离电离)MS / MS测量值相结合,可以精确定位单个N-inking碳水化合物基团,并鉴定N-序列处的焦谷氨酸残基。总站。进一步研究了N-连接的碳水化合物组分,并确定了N-连接的碳水化合物的一般模式。还建立了半胱氨酸78和半胱氨酸24之间的二硫键的存在。与公开的基因序列相比,SBP的全序列表征显示出几个基于菌株的序列差异。

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