Mass Spectrometry Identification

在质谱学研究和支助小组内已经或正在开展蛋白质鉴定方面的各种服务和合作项目，分析了来自31名科学家的约3000个样本，这些科学家代表7个实验室分支机构的23名首席研究人员或核心负责人。 一个大的努力是支持结构生物学核心实验室和Bob Petrovich博士的蛋白质表达功能。MSRSG的作用是在结构生物学核心实验室将材料交给用户之前，在蛋白质水平上确认基因表达。 另一个重要项目是与霍尔实验室合作开展的。在这项研究中，我们确定了与小鼠精子发生有关的潜在调控蛋白。从睾丸提取液中分离出内源性mRNA/蛋白质复合体，用蛋白质质谱仪进行鉴定。这些蛋白质与一个或两个TNP转录本相关。多个蛋白质与TNP转录本有很强的相关性，但在分离肌动蛋白mRNA时信号较弱。这些蛋白在睾丸切片中的表达模式表明，它们在生殖细胞中的适当阶段被检测到，以调节TNP RNA的表达。 其他仍在进行中的未公布项目包括： 质检实验室用MALDI-MS鉴定细菌病原体 BAF的结合伙伴的鉴定(Archer)；TTP(Blackset)；Wnk1(DeMayo)；NTHL1(Doetsch)；Grc3/Las1(Stanley)；Tsen(Stanley)；Smchd1(Shaw)；未定性酯酶(Williams)。 其他需要超过微不足道的资源的项目包括与Jetten、Hu、X.Li和威尔逊实验室进行的工作。 我们还致力于脂类组学和非靶向代谢组学的发展。这两个项目都涉及化合物的鉴定和量化；因此，这些都包括在鉴定项目和量化项目中。 此外，MSRSG还为几个合作者进行蛋白质鉴定，以评估SARS-CoV-2各种蛋白质的表达和纯化。

A variety of service and collaborative projects in protein identification have been or are being carried out within the Mass Spectrometry Research and Support Group with approximately 3000 samples analyzed from 31 scientists representing 23 principal investigators or core heads from 7 laboratory branches. One large effort is in support of the protein expression function of the Structural Biology Core Laboratory and Dr. Bob Petrovich. The Role of the MSRSG is to confirm gene expression at the protein level prior to the Structural Biology Core Laboratory handing materials over to their users. Another significant project was performed in collaboration with the Hall laboratory. In this study, we identified potential regulatory proteins involved in mouse spermatogenesis. Isolated endogenous mRNA/protein complexes from testis extract were identified by mass spectrometry proteins. These proteins associated with one or both Tnp transcripts. Multiple proteins showed strong association with Tnp transcripts but had low signal when Actin mRNA was isolated. Expression patterns in testis sections of these proteins showed that they were detected in germ cells at the appropriate stages to regulate Tnp RNA expression. Other unpublished projects that are still ongoing include: Identification of bacterial pathogens by MALDI-MS with the QA Laboratory. Identification of binding partners of BAF (Archer); TTP (Blackshear); Wnk1 (DeMayo); NTHL1 (Doetsch); Grc3/Las1 (Stanley); TSEN (Stanley); SMCHD1 (Shaw); uncharacterized esterase (Williams). Additional projects that have required more than negligible resources include efforts performed with the Jetten, Hu, X. Li, and Wilson laboratories. We have also put effort into the development of lipidomics and untargeted metabolomics. Both of these projects involve the identification as well as the quantitation of compounds; thus, these are included in both the identification project and the quantitation project. Additionally, the MSRSG performs protein identification to evaluate the expression and purification of various proteins from SARS-CoV-2 for several collaborators.