MASS SPECTROMETRY OF ANTIGENIC LIPOPEPTIDES

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 从一家商业公司获得了一种脂肽的合成版本，该脂肽是最早出现在HIV感染细胞中的化合物之一，并使用来自HIV患者供体的细胞来产生T细胞系。在进一步分析时，发现合成产生的靶向脂肽形式不是纯的，而实际上是混合物，并且进一步地，靶向脂肽不是抗原性的，而是发现两种新的组分是抗原性的。质谱研究表明，新的抗原组分在结构上彼此非常相似，并且具有与预期合成结构相似的结构。在BUSM MS Resource进行的质谱研究包括使用GC/MS、MALDI和ESI Q-o-TOF以及MALDI和ESI-FTMS仪器、已知和未知化合物的酶消化和化学水解。结构现在已经解决;合成类似物已经制备并测试了它们的生物活性。描述这一研究阶段的合作手稿最近出版（I货车Rhijn et al.，CD 1c绕过溶酶体呈递具有12个氨基酸的脂肽抗原。J Exp Med.2009，206，1409-1422.该出版物补充了另一篇报道CD 1a脂肽抗原合成和生物学检测的出版物。(DC Young等人， 由CD 1a呈递的双脱氧分枝杆菌素抗原的合成揭示了T细胞对天然脂肽结构的精细特异性。（J Biol Chem.2009，284，25087-25096）。Madigan女士的论文是探索分支杆菌素-双脱氧粘杆菌素途径的酶途径，以及它们与M。结核病; Costello教授担任她的论文委员会。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. A synthetic version of a lipopeptide that is one of the first compounds to appear in HIV-infected cells was obtained from a commercial company and employed to generate a T cell line, using cells from an HIV patient donor. On further analysis it was found that the synthetically produced version of the targeted lipopeptide was not pure, but in fact a mixture, and further, the targeted lipopeptide was not antigenic, but instead two novel components were found to be antigenic. Mass spectrometric investigations indicate that the new antigenic components are very similar in structure to each other and have structures similar to the intended synthetic structure. Mass spectrometric studies at the BUSM MS Resource included the use of GC/MS, MALDI and ESI Q-o-TOF and MALDI and ESI-FTMS instrumentation, enzymatic digestion and chemical hydrolysis of the known and unknown compounds. The structure has now been solved; synthetic analogs have been prepared and their biological activity has been tested. The collaborative manuscript describing this phase of the research was published recently (I Van Rhijn et al., CD1c bypasses lysosomes to present a lipopeptide antigen with 12 amino acids. J Exp Med. 2009, 206, 1409-1422. This publication complements another that reported on the synthesis and biological testing of CD1a lipopeptide antigens. (DC Young et al., Synthesis of dideoxymycobactin antigens presented by CD1a reveals T cell finespecificity for natural lipopeptide structures. (J Biol Chem. 2009, 284, 25087-25096). Ms. Madigan's thesis is exploring the enzyme pathways that underlie the mycobactin-dideoxymucobactin pathway and their relationship to the variation in lipid profiles in M. tuberculosis; Prof. Costello serves on her thesis committee.