A5028706082- Ubiquitin and proteasome pathways
- Biotin and Related Studies
- Protein Degradation and Inhibitors
- Mitochondrial Function and Pathology
- Neurobiology and Insect Physiology Research
- Multiple Myeloma Research and Treatments
- Click Chemistry and Applications
- Advanced Proteomics Techniques and Applications
- Cellular transport and secretion
- Peptidase Inhibition and Analysis
- Invertebrate Immune Response Mechanisms
- Prostate Cancer Treatment and Research
- Endoplasmic Reticulum Stress and Disease
- Receptor Mechanisms and Signaling
- Glycosylation and Glycoproteins Research
- Plant Molecular Biology Research
- Retinal Development and Disorders
- Toxin Mechanisms and Immunotoxins
- Genetic Neurodegenerative Diseases
- Acute Myeloid Leukemia Research
- Myeloproliferative Neoplasms: Diagnosis and Treatment
- Photochromic and Fluorescence Chemistry
- Hematopoietic Stem Cell Transplantation
- Autophagy in Disease and Therapy
- bioluminescence and chemiluminescence research
Broad Institute
2015-2024
Harvard University
2019-2024
Massachusetts Institute of Technology
2012-2024
University of Michigan–Ann Arbor
2014
Lenalidomide is a drug with clinical efficacy in multiple myeloma and other B cell neoplasms, but its mechanism of action unknown. Using quantitative proteomics, we found that lenalidomide causes selective ubiquitination degradation two lymphoid transcription factors, IKZF1 IKZF3, by the CRBN-CRL4 ubiquitin ligase. IKZF3 are essential factors myeloma. A single amino acid substitution conferred resistance to lenalidomide-induced rescued inhibition growth. Similarly, interleukin-2 production T...
The cytosol-facing membranes of cellular organelles contain proteins that enable signal transduction, regulation morphology and trafficking, protein import export, other specialized processes. Discovery these by traditional biochemical fractionation can be plagued with contaminants loss key components. Using peroxidase-mediated proximity biotinylation, we captured identified endogenous on the outer mitochondrial membrane (OMM) endoplasmic reticulum (ERM) living human fibroblasts. proteomes...
Detection of endogenous ubiquitination sites by mass spectrometry has dramatically improved with the commercialization anti-di-glycine remnant (K-ε-GG) antibodies. Here, we describe a number improvements to K-ε-GG enrichment workflow, including optimized antibody and peptide input requirements, cross-linking, off-line fractionation prior enrichment. This refined practical workflow enables routine identification quantification ∼20,000 distinct in single SILAC experiment using moderate amounts...
Significance Most of the thousands proteins that comprise a human cell have specific subcellular localization patterns essential for their function. “Proximity labeling” (PL) is method mapping endogenous cellular on proteome-wide scale. To improve specificity and versatility PL, we developed split-TurboID, promiscuous biotinylating enzyme split into two inactive fragments. The fragments are coexpressed in cells brought together by drug, protein–protein interaction, or organelle contact to...
Mutant protein in tumors hits the DEK Cancer genome sequencing projects have uncovered a multitude of mutations human tumors. Understanding whether and how these contribute to tumor development progression could ultimately lead new therapies. Theurillat et al. studied product gene that is recurrently mutated prostate cancer. Normally this helps attach biochemical tag cellular proteins marks them for degradation. The work shows tumor-associated mutant loses tagging ability, which results...
Introduction of antibodies specific for acetylated lysine has significantly improved the detection endogenous acetylation sites by mass spectrometry. Here, we describe a new, commercially available mixture anti-lysine (Kac) and show its utility in-depth profiling acetylome. Specifically, seven complementary monoclones with high specificity Kac were combined into final anti-Kac reagent which results in at least twofold increase identification peptides over commonly used antibody. We outline...
Abstract Protein ubiquitylation is involved in a plethora of cellular processes. While antibodies directed at ubiquitin remnants (K-ɛ-GG) have improved the ability to monitor using mass spectrometry, methods for highly multiplexed measurement tissues and primary cells sub-milligram amounts sample remains challenge. Here, we present sensitive, rapid protocol termed UbiFast quantifying ~10,000 sites from as little 500 μg peptide per or tissue TMT10plex ca. 5 h. High-field Asymmetric Waveform...
In developing brains, axons exhibit remarkable precision in selecting synaptic partners among many non-partner cells. Evolutionarily conserved teneurins are transmembrane proteins that instruct partner matching. However, how intracellular signaling pathways execute teneurins' functions is unclear. Here, we use situ proximity labeling to obtain the interactome of a teneurin (Ten-m) Drosophila brain. Genetic interaction studies using quantitative matching assays both olfactory receptor neurons...
While a mutation in C9ORF72 is the most common genetic contributor to amyotrophic lateral sclerosis (ALS), much remains be learned concerning function of protein normally encoded at this locus. To elaborate further on functions for C9ORF72, we used quantitative mass spectrometry-based proteomics identify interacting proteins motor neurons and found that its long isoform complexes with stabilizes SMCR8, which enables interaction WDR41. study organismal cellular tripartite complex, generated...
Transcription factors specify the fate and connectivity of developing neurons. We investigate how a lineage-specific transcription factor, Acj6, controls precise dendrite targeting Drosophila olfactory projection neurons (PNs) by regulating expression cell-surface proteins. Quantitative proteomic profiling wild-type acj6 mutant PNs in intact brains, proteome-informed genetic screen identified PN surface proteins that execute Acj6-regulated wiring decisions. These include canonical cell...
Abstract Driver genes with a mutually exclusive mutation pattern across tumor genomes are thought to have overlapping roles in tumorigenesis. In contrast, we show here that prostate cancer driver alterations involving the ERG transcription factor and ubiquitin ligase adaptor SPOP synthetic sick. At molecular level, incompatible pathways driven by opposing functions SPOP. upregulates wild type dampen androgen receptor (AR) signaling sustain activity through degradation of bromodomain histone...
Abstract Genomic analysis of lung adenocarcinomas has revealed that the MGA gene, which encodes a heterodimeric partner MYC-interacting protein MAX, is significantly mutated or deleted in adenocarcinomas. Most mutations are loss function for MGA, suggesting may act as tumor suppressor. Here, we characterize both molecular and cellular role illustrate its functional relevance MYC pathway. Although interact with same binding partner, recognize E-box DNA motif, show appears to be antagonistic...