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| Open AccessCathepsin D deficiency in mammary epithelium transiently stalls breast cancer by interference with mTORC1 signaling
The lysosomal aspartic protease Cathepsin D (CTSD) is associated with breast cancer progression. Here the authors show that selective inactivation of CTSD in mammary epithelium delays tumor onset due to impaired mTORC1 signaling, but resumes malignant growth due to compensatory oncogenic pathways
- Stephanie Ketterer
- , Julia Mitschke
- & Thomas Reinheckel
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Article
| Open AccessCrystallographic and electrophilic fragment screening of the SARS-CoV-2 main protease
The SARS-CoV-2 main protease is an important target for the development of COVID-19 therapeutics. Here, the authors combine X-ray crystallography and mass spectrometry and performed a large scale fragment screening campaign, which yielded 96 liganded structures of this essential viral protein that are of interest for further drug development efforts.
- Alice Douangamath
- , Daren Fearon
- & Martin A. Walsh
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Article
| Open AccessStructural basis of mammalian mucin processing by the human gut O-glycopeptidase OgpA from Akkermansia muciniphila
OgpA is an O-glycopeptidase from Akkermansia muciniphila, a mucin-degrading bacterium commonly found in the human gut. A thorough characterization of OgpA, including crystal structures in complex with substrate or product, reveals molecular basis of O-glycan recognition and enzyme specificity.
- Beatriz Trastoy
- , Andreas Naegeli
- & Marcelo E. Guerin
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| Open AccessBoth Boceprevir and GC376 efficaciously inhibit SARS-CoV-2 by targeting its main protease
Coronavirus main protease is essential for viral polyprotein processing and maturation. Here Fu et al. report efficient inhibition of SARS-CoV-2 replication using two inhibitors - Boceprevir and GC376 - targeting the active site of the main viral protease.
- Lifeng Fu
- , Fei Ye
- & George Fu Gao
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| Open AccessFeline coronavirus drug inhibits the main protease of SARS-CoV-2 and blocks virus replication
Coronavirus main protease is essential for viral polyprotein processing and replication. Here Vuong et al. report efficient inhibition of SARS-CoV-2 replication by the dipeptide-based protease inhibitor GC376 and its parent GC373, which were originally used to treat feline coronavirus infection.
- Wayne Vuong
- , Muhammad Bashir Khan
- & M. Joanne Lemieux
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Article
| Open AccessIntramolecular chaperone-mediated secretion of an Rhs effector toxin by a type VI secretion system
Bacterial Rhs proteins with toxic domains are often secreted by type VI secretion systems. Here, the authors show that one of these proteins self-cleaves into three fragments, with the Rhs core and the N-terminal domain facilitating secretion and function of the C-terminal toxic domain.
- Tong-Tong Pei
- , Hao Li
- & Tao G. Dong
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Article
| Open AccessA bifunctional asparaginyl endopeptidase efficiently catalyzes both cleavage and cyclization of cyclic trypsin inhibitors
Asparaginyl endopeptidases (AEPs) catalyze the cyclization step during the biosynthesis of cyclic peptides in plants. Here, the authors report a recombinantly produced AEP that catalyzes the backbone cyclization of a linear cyclotide precursor and an engineered analog with high efficiency and in a pH-dependent manner.
- Junqiao Du
- , Kuok Yap
- & David J. Craik
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Article
| Open AccessFAM111A protects replication forks from protein obstacles via its trypsin-like domain
DNA-protein crosslinks represent obstacles on genomic DNA that can hamper progression of replication forks. Here, the authors reveal that FAM111A, a PCNA-interacting protein, plays part in mitigating the effect of protein obstacles on replication forks.
- Yusuke Kojima
- , Yuka Machida
- & Yuichi J. Machida
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Article
| Open AccessCohesin cleavage by separase is enhanced by a substrate motif distinct from the cleavage site
Prior to anaphase, securin binds separase and thereby prevents cohesin cleavage. Here, the authors develop a method to produce active securin-free separase, identify a docking motif in cohesin that promotes cleavage, and show that securin interferes with this interaction.
- Laura E. Rosen
- , Joseph E. Klebba
- & David O. Morgan
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Article
| Open AccessCrystal structure and substrate-induced activation of ADAMTS13
The plasma metalloprotease ADAMTS13 regulates the platelet-tethering function of von Willebrand factor (VWF) in a shear-dependent manner. Here the authors present the ADAMTS13 crystal structure of the 70kDa N-terminal metalloprotease to spacer domains, and using kinetic measurements they identify a substrate binding induced allosteric mechanism for ADAMTS13, where VWF functions both as an activating cofactor and substrate.
- Anastasis Petri
- , Hyo Jung Kim
- & James T. B. Crawley
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Article
| Open AccessAβ34 is a BACE1-derived degradation intermediate associated with amyloid clearance and Alzheimer’s disease progression
Aβ34 is generated from degradation of Aβ40 and Aβ42 by β-secretase. Here, the authors show that Aβ34 is a marker for amyloid clearance and is elevated in the CSF of patients that go on to convert from mild cognitive impairment to Alzheimer’s disease, suggesting it may be a useful biomarker.
- Filip Liebsch
- , Luka Kulic
- & Gerhard Multhaup
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| Open AccessUsp7 regulates Hippo pathway through deubiquitinating the transcriptional coactivator Yorkie
Hippo signaling leads to the phosphorylation of the key transcriptional effector, Yap/Yki, although how Yap/Yki stability is regulated has remained unclear. Here, Sun et al. identify HAUSP/Usp7 as a conserved and clinically relevant regulator of the Hippo pathway that increases Yap/Yki stability.
- Xiaohan Sun
- , Yan Ding
- & Zizhang Zhou
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Article
| Open AccessLysosomal protease deficiency or substrate overload induces an oxidative-stress mediated STAT3-dependent pathway of lysosomal homeostasis
How cells regulate their lysosomal proteolytic capacity is only partly understood. Here, the authors show that lysosomal protease deficiency or substrate overload induces lysosomal stress leading to activation of a STAT3-dependent, TFEB-independent pathway of lysosomal hydrolase expression.
- Jonathan Martínez-Fábregas
- , Alan Prescott
- & Colin Watts
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| Open AccessMapping protein selectivity landscapes using multi-target selective screening and next-generation sequencing of combinatorial libraries
Characterizing the binding selectivity landscape of interacting proteins is crucial in protein engineering. Here the authors use multi-target selective library screening and in silico next-generation sequencing to map the binding landscape of proteins and produce improved proteases inhibitors.
- Si Naftaly
- , Itay Cohen
- & Niv Papo
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Article
| Open AccessIntriguing role of water in protein-ligand binding studied by neutron crystallography on trypsin complexes
Trypsin is a serine protease. Here the authors present the high resolution X-ray and neutron diffraction structures of uncomplexed and inhibitor bound trypsin that provide insights into the geometry of H-bonds in the active site of the enzyme and molecular dynamics simulations reveal the kinetics of ligand binding induced desolvation.
- Johannes Schiebel
- , Roberto Gaspari
- & Gerhard Klebe
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Article
| Open AccessC-terminal truncation of IFN-γ inhibits proinflammatory macrophage responses and is deficient in autoimmune disease
IFN-γ is central in inflammatory pathogenesis, response to infection and autoimmune diseases. Here the authors show that MMP12 expression is reduced in patients with SLE and that MMP12 post-translationally truncates IFN-y, inhibiting its function and affecting pathogenesis of mouse models of peritonitis, SLE and rheumatoid arthritis.
- Antoine Dufour
- , Caroline L. Bellac
- & Christopher M. Overall
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| Open AccessStructural mechanism for nucleotide-driven remodeling of the AAA-ATPase unfoldase in the activated human 26S proteasome
The 26S proteasome consists of a core particle that is capped at each side by a regulatory particle. Here the authors present cryo-EM structures of the activated human 26S proteasome holoenzyme in three alternative open-gate states, which provides mechanistic insights into gate opening and dynamic remodeling of the substrate–translocation pathway.
- Yanan Zhu
- , Wei Li Wang
- & Youdong Mao
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| Open AccessThe beta secretase BACE1 regulates the expression of insulin receptor in the liver
A soluble form of insulin receptor in human plasma has been previously reported. Here the authors demonstrate that insulin receptor is cleaved by BACE1 that can regulate biological active insulin receptor levels in a glucose concentration-dependent manner, both in physiological and diabetic conditions.
- Paul J. Meakin
- , Anna Mezzapesa
- & Franck Peiretti
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| Open AccessReactive-site-centric chemoproteomics identifies a distinct class of deubiquitinase enzymes
Deubiquitinases are proteases that cleave after the C-terminus of ubiquitin to hydrolyze ubiquitin chains and cleave ubiquitin from substrates. Here the authors describe a reactive-site-centric chemoproteomics approach to studying deubiquitinase activity, and expand the repertoire of known deubiquitinases.
- David S. Hewings
- , Johanna Heideker
- & Ingrid E. Wertz
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Article
| Open AccessA common mechanism of proteasome impairment by neurodegenerative disease-associated oligomers
Disruption of the ubiquitin proteasome system (UPS) is often associated with neurodegenerative diseases. Here the authors demonstrate the existence of a general mechanism of proteasomal impairment triggered by a specific protein oligomer structure, irrespective of its protein constituent.
- Tiffany A. Thibaudeau
- , Raymond T. Anderson
- & David M. Smith
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| Open AccessThe protease GtgE from Salmonella exclusively targets inactive Rab GTPases
The bacterial protease GtgE is involved in the establishment of Salmonellosis. Here the authors provide a structural and biochemical analysis of GtgE that sheds light on the molecular mechanisms of reprogramming infected host cells via site-specific proteolytic cleavage of the vesicular trafficking regulator Rab32.
- Rudolf Wachtel
- , Bastian Bräuning
- & Aymelt Itzen
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| Open AccessThe Apaf-1 apoptosome induces formation of caspase-9 homo- and heterodimers with distinct activities
Apoptotic initiator caspases are thought to be activated through homodimerization but this remains controversial. Here the authors demonstrate that caspase-9 can adopt two distinct conformations within the Apaf-1 apoptosome, each with distinct properties that contribute to the overall function of the complex.
- Chu-Chiao Wu
- , Sunhee Lee
- & Shawn B. Bratton
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| Open AccessStructure of the NS2B-NS3 protease from Zika virus after self-cleavage
The proteases of flaviviruses are promising targets for development of specific antiviral drugs. Here, the authors report a high resolution crystal structure of the NS2B-NS3 protease of Zika virus that provides insight into substrate and inhibitor binding.
- Wint Wint Phoo
- , Yan Li
- & Dahai Luo
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Article
| Open AccessEngineered AAA+ proteases reveal principles of proteolysis at the mitochondrial inner membrane
Human YME1L is a membrane-anchored AAA+ protease that maintains proteostasis in the mitochondrial inner membrane and intermembrane space. Here the authors probe the substrate-binding and degradation activities of YME1L and suggest the existence of sequence-specific degradation signals in mitochondrial proteostasis.
- Hui Shi
- , Anthony J. Rampello
- & Steven E. Glynn
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| Open AccessIrreversible inhibitors of the 3C protease of Coxsackie virus through templated assembly of protein-binding fragments
Molecular fragments are useful tools in drug-discovery but they might be hard to identify due to their weak affinity to the targets. Here, the authors use a protein-templated assembly to design high affinity inhibitors of Coxsackie virus 3C protease, a pharmacological target against enteroviral infections.
- Daniel Becker
- , Zuzanna Kaczmarska
- & Jörg Rademann
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| Open AccessPhosphatidylserine exposure is required for ADAM17 sheddase function
ADAM17 is a member of the ‘Disintegrin and Metalloproteinase’ family of proteases, that cleaves transmembrane substrates from the surfaces of cells. Here the authors show that surface exposure of phosphatidylserine is required for ADAM17 sheddase activity, possibly by directing the protease to its substrates.
- Anselm Sommer
- , Felix Kordowski
- & Karina Reiss
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| Open AccessThe unconventional myosin CRINKLED and its mammalian orthologue MYO7A regulate caspases in their signalling roles
In addition to their role in apoptosis, caspases are also involved in mediating non-apoptotic events. Here the authors show that the Drosophilamyosin family member CRINKLED and its mammalian counterpart act as substrate adaptor that facilitate caspase-mediated cleavage and localised kinase activity.
- Mariam H. Orme
- , Gianmaria Liccardi
- & Pascal Meier
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| Open AccessNanocaged enzymes with enhanced catalytic activity and increased stability against protease digestion
Cells compartmentalize enzymes for enhanced efficiency of their metabolic pathways. Here, the authors describe a self-assembly approach to construct DNA nanocaged enzymes for enhancing catalytic activity and stability, and observe an inversed correlation between the protein size and the activity enhancement.
- Zhao Zhao
- , Jinglin Fu
- & Hao Yan
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Article
| Open AccessMolecular sled is an eleven-amino acid vehicle facilitating biochemical interactions via sliding components along DNA
Extremely compact environments can inhibit protein interactions by preventing 3-dimensional diffusion. Here the authors show that an 11-amino acid long peptide can function as a ‘molecular sled’, able to transport cargo linearly along DNA to enable interactions.
- Walter F. Mangel
- , William J. McGrath
- & Paul C. Blainey
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Article
| Open AccessA role for Separase in telomere protection
Drosophila telomeres are elongated by transposition of specialized retroelements rather than telomerase activity. Here, the authors show that Separase is enriched at Drosophila telomeres and loss of Sse, the gene encoding Separase, leads to telomere defects, suggesting a role for Separase in telomere protection.
- Francesca Cipressa
- , Patrizia Morciano
- & Giovanni Cenci
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| Open AccessCatalytic site inhibition of insulin-degrading enzyme by a small molecule induces glucose intolerance in mice
Inhibiting insulin-degrading enzyme (IDE) has been proposed as a potential therapeutic strategy for the treatment of patients with diabetes. Here, the authors develop a novel IDE inhibitor but find that, surprisingly, IDE inhibition has negative effects on glucose tolerance in mice.
- Rebecca Deprez-Poulain
- , Nathalie Hennuyer
- & Benoit Deprez
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Redefining the concept of protease-activated receptors: cathepsin S evokes itch via activation of Mrgprs
Sensory neurons that mediate histamine-independent itch express Mas-related G protein coupled receptors (Mrgprs). Here, Reddy et al.show that the cysteine protease cathepsin S cleaves and activates MrgpcrC11 without the generation of a tethered ligand, in contrast to other protease activated receptors.
- Vemuri B. Reddy
- , Shuohao Sun
- & Ethan A. Lerner
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| Open AccessEarly emergence of Yersinia pestis as a severe respiratory pathogen
Yersinia pestis, which evolved from a gastrointestinal pathogen, causes pneumonic and bubonic plague. Here Zimbler et al. show that the gain of a single protein enabled Y. pestisto first cause pneumonic plague, and one amino-acid change in the same protein then allowed the bacteria to efficiently cause bubonic plague.
- Daniel L. Zimbler
- , Jay A. Schroeder
- & Wyndham W. Lathem
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| Open AccessSystem-wide identification of wild-type SUMO-2 conjugation sites
Tryptic digestion of SUMOylated proteins generates large peptides, rendering proteomic characterisation of this post-translational modification particularly challenging unless mutant SUMO is used. Hendriks et al.present a method that allows the quantitative identification of wild-type SUMO sites.
- Ivo A. Hendriks
- , Rochelle C. D’Souza
- & Alfred C. O. Vertegaal
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Article
| Open Accessγ-secretase directly sheds the survival receptor BCMA from plasma cells
B-cell maturation antigen (BCMA) regulates the survival of B cells and is essential for the maintenance of long-lived plasma cells. Here, the authors show that γ-secretase directly sheds BCMA from the cell surface and therefore regulates the number of plasma cells.
- Sarah A. Laurent
- , Franziska S. Hoffmann
- & Edgar Meinl
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Article |
Autoprocessing of neutrophil elastase near its active site reduces the efficiency of natural and synthetic elastase inhibitors
Elastase secreted by immune cells contributes to various lung diseases; however, elastase inhibitors have mostly failed in the clinic. Here, the authors discover a second, truncated form of elastase, which is the result of autocatalytic cleavage and is not well targeted by current synthetic elastase inhibitors.
- T. Dau
- , R. S. J. Sarker
- & D. E. Jenne
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A system for the continuous directed evolution of proteases rapidly reveals drug-resistance mutations
Phage-assisted continuous evolution (PACE) has the potential to rapidly evolve drug-resistant mutations. Here, Dickinson et al.present a protease PACE system that identifies clinically relevant mutations conferring resistance to protease inhibitors in only a few days of continuous evolution.
- Bryan C. Dickinson
- , Michael S. Packer
- & David R. Liu
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Article |
Out-of-frame start codons prevent translation of truncated nucleo-cytosolic cathepsin L in vivo
The lysosomal protease cathepsin L has been observed in compartments other than endosomes and lysosomes. Here the authors show using knock-in mice that nuclear localization of cathepsin L cannot be caused by N-terminal truncation of procathepsin L as previously hypothesized.
- Martina Tholen
- , Larissa E. Hillebrand
- & Thomas Reinheckel
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Article
| Open AccessHuman CLPP reverts the longevity phenotype of a fungal ClpP deletion strain
The mitochondrial protease CLPP is found in most eukaryotic organisms but its biological role has been unclear. Here Osiewacz and colleagues show that deletion of CLPP extends lifespan of the filamentous fungus Podospora anserina, and that human and fungal CLPP are functionally conserved.
- Fabian Fischer
- , Andrea Weil
- & Heinz D. Osiewacz
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Article
| Open AccessSubstrate docking to γ-secretase allows access of γ-secretase modulators to an allosteric site
γ-Secretase modulators have promise in the treatment of Alzheimer's disease, but their molecular target is uncertain. Here, fluorescence resonance energy transfer is used to determine that the γ-secretase allosteric site is within the γ-secretase complex and that substrate docking is required for modulators to access the site.
- Kengo Uemura
- , Katherine C. Farner
- & Oksana Berezovska