MASS SPECTROMETRY OF DRUGS, NATURAL PRODUCTS, PROTEINS, AND OLIGONUCLEOTIDES

该单位的研究分为从天然来源分离的生物活性微量有机化合物的鉴定和生物聚合物（如肽，蛋白质和寡核苷酸）的质谱分析。与NHLBI生物物理化学实验室保持着非常密切的工作关系和合作。虽然这个小组为NIH的实验室进行了许多常规分析，但尽可能多地强调使用质谱法研究生物活性化合物的结构。今年的重点项目包括：强效抗艾滋病和抗癌化合物的结构鉴定；蛋白质翻译后修饰的鉴定；继续研究强效抗艾滋病毒化合物氰维林（cyanovirin），该药物即将进入临床试验阶段；发展生物聚合物的微尺度LCMS技术，以期大大提高灵敏度；海洋天然产物中异常氨基酸和修饰氨基酸的鉴定及手性测定镰状细胞治疗过程中形成的修饰血红蛋白的鉴定；开发了一种利用氘交换跟踪蛋白质折叠的方法。在这一年中，所有研究的重点都集中在具有多个糖基化位点和许多重叠二硫化物的生物聚合物和大蛋白质上。人们正在尝试用质谱法来定义3-D蛋白质的形状。多种酶消化方案和复杂的软件已经开发出来，以协助研究，这些现在在世界范围内使用。一个新的Q-TOF质谱计已经与其他一些研究所共享，这将极大地扩大生物聚合物的研究潜力。

Research of the unit is split between the identification of biologically active trace organic compounds isolated from natural sources and the mass spectrometry of biopolymers (e.g. peptide, proteins and oligonucleotides). A very close working relationship and collaboration is maintained with the Laboratory of Biophysical Chemistry, NHLBI. While this group runs many routine analyses for laboratories at NIH, as much emphasis as possible is placed on research using mass spectrometry for the structural elucidation of biologically active compounds. This year's key projects include: the structural identification of potent anti-HIV and anti-cancer compounds; the identification of post-translational modifications in proteins; continued research on cyanovirin, a potent anti-HIV compound heading towards clinical trial; development of techniques for micro- scale LCMS of biopolymers with the aim of greatly enhancing sensitivity; the identification and chiral determination of abnormal and modified amino acids as found in marine natural products; the identification of modified hemoglobins formed during treatments for sickle cell; and development of a method to follow protein folding using deuterium exchange. During the year the emphasis of all the research has become biopolymers and large proteins with multiple glycosylation sites and many overlapping disulfides and being examined. Attempts to define 3-D protein shapes using mass spectrometry are being developed. Multiple enzymatic digestion protocols and sophisticated software have been developed to assist the research and these are now in use worldwide. A new Q-TOF mass spectreometer shared with some other institutes has been ordered and this will drastically expand the biopolymer research potential of the group.