MASS SPECTROMETRY OF ANTIGENIC LIPOPEPTIDES

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 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。 描述这一阶段研究的手稿最近发表（I货车Rhijn等人，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份最近发表的关于CD 1a脂肽抗原合成和生物学检测的报告。(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）。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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 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 recent publication 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.)