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 Other unpublished projects that are still ongoing include: Identification of bacterial pathogens by MALDI-MS with the QA Laboratory.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

质谱研究和支持小组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7个实验室分支机构的23名主要研究者或核心负责人的31名科学家的约3000个样本。 一个大的努力是支持结构生物学核心实验室和鲍勃彼得罗维奇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其用户之前，在蛋白质水平上确认基因表达。 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，我们确定了潜在的调控蛋白参与小鼠精子发生。 从睾丸提取物中分离的内源性mRNA/蛋白质复合物通过质谱蛋白质鉴定。 这些蛋白质与一种或两种Tnp转录物相关。多种蛋白质显示与Tnp转录物的强关联，但当分离肌动蛋白mRNA时具有低信号。这些蛋白质在睾丸切片中的表达模式表明，它们在生殖细胞中检测到在适当的阶段调节Tnp RNA表达。 其他尚未公布的项目仍在进行中，包括： QA实验室通过MALDI-MS鉴定细菌病原体。 BAF（Archer）; TTP（黑剪）; Wnk 1（DeMayo）; NTHL 1（Doetsch）; Grc 3/Las 1（Stanley）; TSEN（Stanley）; SMCHD 1（Shaw）;未表征酯酶（威廉姆斯）的结合配偶体的鉴定。 其他项目所需的资源可忽略不计，包括Jetten、Hu、X。李和威尔逊实验室 我们还致力于脂质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定量;因此，这些都包括在鉴定项目和定量项目中。 此外，MSRSG进行蛋白质鉴定，以评估来自SARS-CoV-2的各种蛋白质的表达和纯化。