摘要
目的 采用质谱技术研究许旺细胞源神经营养蛋白 (SDNP)的分子结构。 方法 将纯化后保持活性的SDNP ,用TPCK 胰蛋白酶酶解 ,行激光飞行时间质谱检测 ,测定肽质量指纹质谱 ,以及碎片结构分析测序列 ,然后在相应的质谱数据库中检索。 结果 (1)激光飞行时间质谱测得完整样品中SDNP的分子量 ;(2 )同时测得酶解样品中几十个肽段的质谱图 ,各肽段的分子量亦相应测出 ,检测8个肽段参数 ,通过蛋白质谱数据库MS Fit检索未发现有蛋白 10 0 %吻合 ,Hypotheticalprotein有 5个肽段符合 (吻合率 62 % ) ;(3 )取酶解质谱中的 2 3 0 5Da片段经质谱碎片分析得到若干碎片离子类型图谱 ,从获得的参数通过蛋白质谱数据库MS Tag检索得出该片段的氨基酸序列。 结论 通过质谱检测 ,许旺细胞源神经营养蛋白的相对分子质量是 66× 10 3 ,部分片段的氨基酸序列为EPVKKVTNSRRAKP TKPNGHIAN。
Objective To analyse the molecular structure of Schwann cell derived neurotrophic protein (SDNP) by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF MS) Methods The purity of SDNP was digested byTPCK trypsin and detected by MALDI TOF MS, mass mapping was determined and partial sequences of the protein have neen analyzed by the mass data of fragment ion peaks in the fragmentation analysis and structural TOF MS (FAST) spectra The primary partial structure of SDNP was identified by searching the protein databases Results The integrity SDNP in peak detected by MALDI TOF MS has 66×10 3 MW and higher purity, because no other peaks exists except double charge peak The mass mapping of many peptides was determined and 8 peptides of them have been retrieved in MS Fit database, there is not the same protein in database Hypothetical protein has 5 peptides homology with the sample (62%) FAST spectra of 2305 Da shows the primary partial structure of SDNP after searching the protein MS Tag database Conclusions The molecular weight of SDNP detected by MALDI TOF MS is 66×10 3, The sequence of partial amino acid is EPVKKVTNSRRAKRTKPNGHIAN
出处
《中华显微外科杂志》
CSCD
北大核心
2002年第4期271-273,共3页
Chinese Journal of Microsurgery
基金
中国博士后科学基金 (2 0 0 0 2 3)
广东省卫生厅基金(A2 0 0 0 1 51 )
关键词
许旺细胞
神经营养
蛋白质
质谱
氨基酸序列
Schwann cells
Neurotrophy
Protein
Mass spectrometry
Amino acid sequence
