PROTEIN SEQUENCING BY TANDEM MASS SPECTROMETRY

研究解决蛋白质化学和蛋白质化学中的一些结构问题 继续发展仪器和方法学，以 提出了串联质谱仪进行蛋白质序列分析的方法。 将作出努力；(A)开发用于直接序列的光谱分析 飞摩尔/皮科莫尔分离的疾病诊断蛋白的分析 用二维凝胶从培养细胞和生物体液中提取蛋白质 电泳法，(B)表征结构修饰， 人类晶状体中的蛋白质在白内障形成过程中经历(C)使用 串联质量相结合的激光光交联法 DNA聚合酶I中氨基酸残基的精确定位 在合成进行时的结合域和 脱氧核苷酸三磷酸结合位点的下一个互补 核苷酸被持有，(D)建立两个钙的一级结构 参与脑内神经元分化的结合蛋白，(E)至 肌球蛋白调控轻链亚型的序列分析 在血管平滑肌中维持高血压，(F)至 描述尼古丁的地形或折叠模式 构成配体门控离子通道的乙酰胆碱受体蛋白 在神经肌肉接头，(G)定位9个磷酸化 视觉蛋白中的残留物视紫红质，并确定其顺序 这些残基被磷酸化是理解的第一步 磷酸化在视觉过程调节中的作用 分子水平，(H)建立人的一级结构 参与免疫调节的单核细胞趋化蛋白 对抗原的反应，以及(I)对氨酰基酶和两个 磷酸酪氨酸磷酸酶参与细胞增殖。努力将会 也要减少分析所需的样本量 新获得的270型三重四极杆质谱仪 皮摩尔范围和扩大该仪器的可用质量范围 超过3,000马力。毛细管电泳会与 电喷雾离子三极四极和四极FTMS仪器 促进蛋白质和寡肽样本在线分析的来源 被这一极其强大的技术隔开。倍增的碎片 带电的多肽离子将被研究，并允许 异亮氨酸和亮氨酸残基的区别 碰撞激活解离或表面诱导解离将 探索过了。

Research to solve a number of structural problems in protein chemistry and to continue the development of instrumentation and methodology for the sequence analysis of proteins by tandem mass spectrometry is proposed. Efforts will be made; (a) to develop spectrometry for the direct sequence analysis of disease diagnostic proteins isolated at the femtomole/picomole level from cultured cells and biological fluids by two dimensional gel electrophoresis, (b) to characterize the structural modifications that proteins in the human lens undergo during cataract formation, (c) to use the laser photocrosslinking technique in combination with tandem mass spectrometry to pinpoint amino acid residues in DNA polymerase I involved in both the binding domain at which synthesis proceeds and the deoxynucleotide triphosphate binding site where the next complementary nucleotide is held, (d) to establish the primary structures of two calcium binding proteins involved in neuronal differentiation in the brain, (e) to sequence the regulatory light chain isoforms of myosin involved in maintaining high blood pressure in vascular smooth muscle, (f) to characterize the topography or folding pattern of the nicotinic acetylcholine receptor protein that makes up the ligand gated ion channel at the neuromuscular junction, (g) to locate the nine phosphorylated residues in the visual protein, rhodopsin, and to determine the order in which these residues are phosphorylated as a first step in understanding the role that phosphorylation plays in regulation of the visual process at the molecular level, (h) to establish the primary structures of human monocyte chemoattractant proteins involved in modulating the immune response to antigens, and (i) to sequence aminoacylase and two phosphotyrosyl phosphatases involved in cell proliferation. Efforts will also be made to reduce the amount of sample required for analysis on the newly acquired Model 270 triple quadrupole mass spectrometer to the low picomole range and to extend the usable mass range of this instrument beyond m/z 3,000. Capillary electrophoresis will be interfaced to the triple quadrupole and quadrupole FTMS instruments via electrospray ion sources to facilitate online analysis of protein and oligopeptide samples separated by this extremely powerful technique. Fragmentation of multiply charged peptide ions will be studied and conditions that allow differentiation of leucine and isoleucine residues by the process of collision activated dissociation or surface induced dissociation will be explored.