Mass Spectrometry Identification

质谱鉴定

• 批准号: 10253944
• 负责人: Jason Williams
• 金额: $ 72.1万
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10253944
• 关键词: 2019-nCoV; Actins; Binding; Collaborations; Development; Gene Expression; Genetic Transcription; Germ Cells; Hand; Head; Laboratories; Mass Spectrum Analysis; Messenger RNA; Mus; National Institute of Environmental Health Sciences; Pattern; Principal Investigator; Proteins; Research Personnel; Research Support; Resources; Role; Sampling; Scientist; Services; Signal Transduction; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spermatogenesis; Support Groups; Testis; Transcript; base; esterase; genetic regulatory protein; metabolomics; pathogenic bacteria; protein complex; protein expression; structural biology

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 Other unpublished projects that are still ongoing include: Identification of bacterial pathogens by MALDI-MS with the QA Laboratory.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.

在质谱研究和支持小组中，已经或正在进行了各种蛋白质识别的服务和协作项目，其中大约3000个样本，分析了31位科学家，代表23名主要研究人员或来自7个实验室分支机构的核心负责人。 一项巨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移交给使用者之前确认蛋白质水平上的基因表达。 另一个重要的项目是与霍尔实验室合作进行的。 在这项研究中，我们确定了与小鼠精子发生有关的潜在调节蛋白。 通过质谱蛋白鉴定出睾丸提取物中分离出的内源性mRNA/蛋白质复合物。 这些蛋白质与一个或两个TNP转录本相关。多种蛋白质与TNP转录本显示强大的关联，但是分离肌动蛋白mRNA时的信号较低。这些蛋白质的睾丸切片中的表达模式表明，在适当阶段在生殖细胞中检测到它们以调节TNP RNA表达。 其他仍在进行的未发表的项目包括： 通过质量检查实验室对MALDI-MS鉴定细菌病原体。 识别BAF（Archer）的约束伙伴； TTP（黑皮）; wnk1（demayo）; nthl1（doetsch）; GRC3/LAS1（Stanley）; Tsen（斯坦利）； SMCHD1（Shaw）;未表征的酯酶（Williams）。 除了可忽略的资源所需的其他项目还包括与Jetten，Hu，X. Li和Wilson Laboratories所做的努力。 我们还为脂肪组学和非靶向代谢组学的发展付出了努力。 这两个项目都涉及化合物的识别和定量。因此，这些都包含在标识项目和定量项目中。 此外，MSRSG执行蛋白质鉴定，以评估几个合作者的SARS-COV-2各种蛋白质的表达和纯化。