A5027450456- Ubiquitin and proteasome pathways
- Microtubule and mitosis dynamics
- Autophagy in Disease and Therapy
- Genomics and Chromatin Dynamics
- DNA Repair Mechanisms
- Peptidase Inhibition and Analysis
- Histone Deacetylase Inhibitors Research
- Chromosomal and Genetic Variations
- Lipid metabolism and biosynthesis
- Endoplasmic Reticulum Stress and Disease
- Photosynthetic Processes and Mechanisms
- Cancer-related Molecular Pathways
- Genetics and Neurodevelopmental Disorders
- DNA and Nucleic Acid Chemistry
- CRISPR and Genetic Engineering
- Advanced Fluorescence Microscopy Techniques
- Protein Structure and Dynamics
- PARP inhibition in cancer therapy
- Cancer-related gene regulation
- Mitochondrial Function and Pathology
- Organizational Management and Leadership
- Advanced biosensing and bioanalysis techniques
- Cannabis and Cannabinoid Research
- Calcium signaling and nucleotide metabolism
- Cellular transport and secretion
Max Planck Institute for Multidisciplinary Sciences
2022-2023
Max Planck Institute for Dynamics and Self-Organization
2022-2023
Max Planck Institute for Biophysical Chemistry
2021
Max Planck Institute of Molecular Physiology
2015-2018
Max Planck Society
2015
The Netherlands Cancer Institute
2010-2014
Eindhoven University of Technology
2006
The fusion of different protein domains via peptide linkers is a powerful, modular approach to obtain proteins with new functions. A detailed understanding the conformational behavior important for applications such as fluorescence resonance energy transfer (FRET)-based sensor and multidomain involved in multivalent interactions. To investigate flexible glycine- serine-containing linkers, we constructed series enhanced cyan yellow fluorescent (ECFP-linker-EYFP) which linker length was...
Autophagy is a conserved intracellular degradation pathway that generates de novo double-membrane autophagosomes to target wide range of material for lysosomal degradation. In multicellular organisms, autophagy initiation requires the timely assembly contact site between ER and nascent autophagosome. Here, we report in vitro reconstitution full-length seven-subunit human supercomplex built on core complex ATG13-101 ATG9. Assembly this rare ability ATG13 ATG101 switch distinct folds. The slow...
Abstract Ubiquitin-specific protease USP4 is emerging as an important regulator of cellular pathways, including the TGF-β response, NF-κB signalling and splicing, with possible roles in cancer. Here we show that has its catalytic triad arranged a productive conformation. Nevertheless, it requires N-terminal DUSP–Ubl domain to achieve full turnover. Pre-steady-state kinetics measurements reveal activity strongly inhibited by slow dissociation ubiquitin after substrate hydrolysis. The able...
The approximately thirty core subunits of kinetochores assemble on centromeric chromatin containing the histone H3 variant CENP-A and connect chromosomes with spindle microtubules. proximal 16-subunit CCAN (constitutive centromere associated network) creates a mechanically stable bridge between kinetochore's microtubule-binding machinery, 10-subunit KMN assembly. Here, we reconstituted stoichiometric 11-subunit human that forms when CENP-OPQUR complex binds to joint interface CENP-HIKM...
We demonstrate that oxime ligation is an efficient, straightforward, and generally applicable strategy for generating nonhydrolyzable ubiquitin (Ub)-isopeptide isosteres. synthesized K48- K63-linked Ub-isopeptide isosteres to investigate the selectivity of deubiquitinating enzymes specific linkages employing surface plasmon resonance spectroscopy. The results indicate specifically recognize local peptide sequence flanking Ub-branched lysine residues in target proteins. described allows...
Accurate chromosome segregation during cell division is crucial for propagating life and protects from cellular transformation. The SKAP:Astrin heterodimer localizes to spindle microtubules mature microtubule-kinetochore attachments mitosis. Depletion of either subunit disrupts structure destabilizes kinetochore-microtubule attachments. Here, we identify molecular requirements the inter-subunit interaction SKAP Astrin, discuss their kinetochore recruitment. We also characterize a...
MAD2L2 (REV7) plays an important role in DNA double-strand break repair. As a member of the shieldin complex, consisting MAD2L2, SHLD1, SHLD2 and SHLD3, it controls repair pathway choice by counteracting end-resection. Here we investigated requirements for complex assembly activity. Besides dimerization-surface, HORMA-domain protein has extraordinary ability to wrap its C-terminus around likely creating very stable complex. We show that appropriate function within requires dimerization,...
Scientific Report18 March 2011Open Access Ubiquitin-specific protease 4 is inhibited by its ubiquitin-like domain Mark P A Luna-Vargas Division of Biochemistry and Center for Biomedical Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, Search more papers this author Alex C Faesen Willem J van Dijk Michael Rape Department Molecular Cell Biology, University California at Berkeley, California, 94720 USA Alexander Fish Titia K Sixma Corresponding Author Information...
The spindle assembly checkpoint (SAC) prevents premature sister chromatid separation during mitosis. Phosphorylation of unattached kinetochores by the Mps1 kinase promotes recruitment SAC machinery that catalyzes effector mitotic complex (MCC). protein Bub3 is a phospho-amino acid adaptor forms structurally related stable complexes with functionally distinct paralogs named Bub1 and BubR1. A short motif ("loop") Bub1, but not equivalent loop BubR1, enhances binding to kinetochore...
Large supramolecular protein complexes, such as the molecular machinery involved in gene regulation, cell signaling, or division, are key all fundamental processes of life. Detailed elucidation structure and dynamics complexes can be achieved by reverse-engineering parts order to probe their interactions with distinctive binding partners vitro. The exploitation DNA nanostructures mimic partially assembled which presence state two more proteins decisive for additional building blocks is...
The Shieldin complex represses end resection at DNA double-strand breaks (DSBs) and thereby serves as a pro-non homologous joining (NHEJ) factor. molecular details of the assembly its recruitment to DSBs are unclear. contains two REV7 molecules, which have rare ability slowly switch between multiple distinct native states could dynamically control Shieldin. Here, we report identification promiscuous binding domain in SHLD3. At N-terminus, SHLD3 interacts with dimer molecules. We show that...
Autophagosome biogenesis requires the timely assembly of a complex machinery. However, it is unclear how putative stable super-complex assembled and disassembled, different parts cooperate to perform its overall function. To shed light on these questions, we embarked an effort biochemically reconstitute human initiation complexes in vitro with purified full-length components. This approach revealed that all subcomplexes together form interaction platform consisting ATG9A, ATG13 ATG101. An...
Abstract The Shieldin complex represses end resection at DNA double-strand breaks (DSBs) and thereby serves as a pro-non homologous joining (NHEJ) factor in the G1 phase of cell cycle. Its components SHLD1, SHLD2, SHLD3 REV7 are recruited hierarchical fashion. localize first to DSBs, while subsequently SHLD2 is only known binding protein complex. molecular details initial recruitment REV7, subsequent assembly on DSBs unclear. Here, we report identification promiscuous domain C-terminal half...
Abstract Autophagy is a conserved intracellular degradation pathway that uses de novo doublemembrane vesicle (autophagosome) formation to target wide range of cytoplasmic material for lysosomal degradation. In multicellular organisms, autophagy initiation requires the timely assembly contact site between ER and nascent autophagosome. Here, we report in vitro reconstitution full-length sevensubunit human supercomplex found at its core ATG13-101 transmembrane protein ATG9. Assembly this...