Enzymes

  • Article
    | Open Access

    During embryonic development, neural progenitor cells undergo numerous cell divisions. Here, the authors show that ABHD4-mediated developmental anoikis distinguishes the physiological delamination and the pathological detachment of progenitor cells with relevance to fetal alcohol-induced apoptosis.

    • Zsófia I. László
    • , Zsolt Lele
    •  & István Katona
  • Article
    | Open Access

    A cell free or synthetic biochemistry approach offers a way to circumvent the many constraints of living cells. Here, the authors demonstrate, via enzyme and process enhancements, the production of isobutanol with the metrics exceeding highly developed ethanol fermentation.

    • Saken Sherkhanov
    • , Tyler P. Korman
    •  & James U. Bowie
  • Article
    | Open Access

    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
  • Article
    | Open Access

    Impaired oligodendrocyte (OL) differentiation and remyelination after myelin damage in multiple sclerosis is associated with neurodegeneration. The authors show that Gsta4 is expressed during adult OL differentiation and identify it as a regulator of OL differentiation, survival, and remyelination.

    • Karl E. Carlström
    • , Keying Zhu
    •  & Fredrik Piehl
  • Article
    | Open Access

    2-oxoglutarate links mitochondrial metabolism to oxygen sensing, gene transcription and epigenetic modifications. Here, the authors describe a mutagenesis screen to find genes that control 2-oxoglutarate levels, and identify ABHD11 as a mitochondrial regulator of 2-oxoglutarate abundance.

    • Peter S. J. Bailey
    • , Brian M. Ortmann
    •  & James A. Nathan
  • Article
    | Open Access

    Plasmodium infection activates signaling pathways in a-nucleated erythrocytes. Here, Adderley et al. use a comprehensive antibody microarray to show that infection extensively modulates host cell signalling and that the host receptor tyrosine kinase c-MET supports Plasmodium falciparum proliferation.

    • Jack D. Adderley
    • , Simona John von Freyend
    •  & Christian Doerig
  • Article
    | Open Access

    Terpene cyclases catalyze the formation of diverse hydrocarbon scaffolds found in terpenoids. Here, the authors report the cryptic function of class I terpene cyclases as aromatic prenyltransferases and the universality of this cryptic feature is confirmed using enzymes from different sources.

    • Haibing He
    • , Guangkai Bian
    •  & Tiangang Liu
  • Article
    | Open Access

    Monooxygenases catalyse the hydroxylation of C-H bonds using oxygen as a co-substrate, which, in turn, is unavailable for anaerobic bacteria. Here, the authors report a three-step reaction cascade involving two hydroxylases and one dehydratase which hydroxylate the C26 methyl group of cholesterol with water as a co-substrate.

    • Christian Jacoby
    • , Sascha Ferlaino
    •  & Matthias Boll
  • Article
    | Open Access

    Specific non-structural proteins (nsp) of SARS coronaviruses are involved in methylation of virally encoded mRNAs to mimic cellular mRNAs for protection against host innate immune restriction. Here, the authors present a high resolution structure of SARS-CoV-2 nsp16/nsp10 ternary complex in the presence of cognate RNA substrate analogue and methyl donor, S-adenosyl methionine, revealing unique ligand-binding sites that may represent alternative targets for antiviral development.

    • Thiruselvam Viswanathan
    • , Shailee Arya
    •  & Yogesh K. Gupta
  • Article
    | Open Access

    The cryo-EM structure of Mycobacterium smegmatis arabinosyltransferase B EmbB involved in mycobacterial cell wall biosynthesis provides insights into the substrate binding and reaction mechanism. Mapping of the ethambutol resistance associated mutations onto the structure suggests the location of the drug binding site.

    • Yong Zi Tan
    • , José Rodrigues
    •  & Filippo Mancia
  • Article
    | Open Access

    Receptor-linked protein tyrosine phosphatases (RPTPs) usually contain an active membrane proximal domain and an inactive pseudophosphatase domain. Here, the authors characterize an RPTP with two pseudophosphatase domains, providing evidence that it may act as a decoy receptor for substrate sequestration.

    • Iain M. Hay
    • , Gareth W. Fearnley
    •  & Janet E. Deane
  • Article
    | Open Access

    Chemical tools to monitor drug-target engagement of endogenous enzymes are essential for preclinical target validation. Here, the authors present a chemical genetics strategy to study target engagement of endogenous kinases, achieving specific labeling and inactivation of FES kinase to provide insights into FES’ role in neutrophil phagocytosis.

    • Tom van der Wel
    • , Riet Hilhorst
    •  & Mario van der Stelt
  • Article
    | Open Access

    The SARS-CoV-2 3CL main protease (3CL Mpro) is a chymotrypsin-like protease that facilitates the production of non-structural proteins, which are essential for viral replication and is therefore of great interest as a drug target. Here, the authors present the 2.30 Å room temperature crystal structure of ligand-free 3CL Mpro and compare it with the earlier determined low-temperature ligand-free and inhibitor-bound crystal structures.

    • Daniel W. Kneller
    • , Gwyndalyn Phillips
    •  & Andrey Kovalevsky
  • Article
    | Open Access

    Flavins and deazaflavins are well suited for photoredox processes but their application in photoreductions is challenging. Here, the authors provide direct evidence of the high reductive power of excited deazaflavin semiquinones and their application in catalytic photodehalogenations.

    • Andreas Graml
    • , Tomáš Neveselý
    •  & Burkhard König
  • Article
    | Open Access

    Complex polyketides are usually produced by microbes, whereas the origin of polyketides found in animals remained unknown. This study shows that sacoglossan animals, such as sea slugs, employ fatty acid synthase-like proteins to produce microbe-like polyketides.

    • Joshua P. Torres
    • , Zhenjian Lin
    •  & Eric W. Schmidt
  • Article
    | Open Access

    Enzymes from organisms inhabiting cold environments (psychrophiles) have adapted to catalyzing chemical reactions at near freezing temperatures. Here – using molecular dynamics simulations – the authors analyze cold adaptation of psychrophilic α-amylase and provide the structural basis for its low anomalous temperature optimum: the increased mobility of a surface loop involved in substrate interaction.

    • Jaka Sočan
    • , Miha Purg
    •  & Johan Åqvist
  • Article
    | Open Access

    The number of usable light-responsive enzymes is limited, despite the potential biotechnological applications. Here, the authors report a flavoprotein monooxygenase which is controllable by blue light illumination, and propose a mechanism involving protein-mediated radical photoreduction of FAD via a semiquinone intermediate.

    • Simon Ernst
    • , Stefano Rovida
    •  & Steffen L. Drees
  • Article
    | Open Access

    Protein arginine methyltransferases (PRMTs) are increasingly recognized as potential therapeutic targets but PRMT7 remains an understudied member of this enzyme family. Here, the authors develop a chemical probe for PRMT7 and apply it to elucidate the role of PRMT7 in the cellular stress response.

    • Magdalena M. Szewczyk
    • , Yoshinori Ishikawa
    •  & Dalia Barsyte-Lovejoy
  • Article
    | Open Access

    The DNA methyltransferase DNMT3A plays an important role in establishing the DNA methylation patterns during development and deregulation of DNMT3A is associated with hematological cancers, with the R882H mutation the most frequently occurring DNMT3A missense mutation in acute myeloid leukemia. Here, the authors present the crystal structures of wild-type and R882H DNMT3A in complex with different DNA substrates and explain why the R882H mutation compromises the enzymatic activity of DNMT3A.

    • Hiwot Anteneh
    • , Jian Fang
    •  & Jikui Song
  • Article
    | Open Access

    Gene rv3722c is essential for in vitro growth of Mycobacterium tuberculosis, but its function is unclear. Here, Jansen et al. show that Rv3722c is the primary aspartate aminotransferase of this pathogen, mediates nitrogen distribution, and is important for virulence during infection of macrophages and mice.

    • Robert S. Jansen
    • , Lungelo Mandyoli
    •  & Kyu Y. Rhee
  • Article
    | Open Access

    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
  • Article
    | Open Access

    Mfd recognizes stalled transcriptional complexes at sites of lesions and recruits the nucleotide excision repair proteins (UvrAB) to the site. Here the authors use live cell imaging in E. coli to demonstrate that coordinated ATP hydrolysis by UvrA and loading of UvrB on DNA facilitate the dissociation of Mfd from the handoff complex.

    • Han Ngoc Ho
    • , Antoine M. van Oijen
    •  & Harshad Ghodke
  • Article
    | Open Access

    O-alkylation of carboxylates by alkyl halides has only been observed transiently in enzymatic processes. Here, the authors show a carboxylate alkylating enzyme, BrtB, that catalyzes C-O bond formation between free fatty acids and secondary alkyl chlorides.

    • João P. A. Reis
    • , Sandra A. C. Figueiredo
    •  & Pedro N. Leão
  • Article
    | Open Access

    Acetaldehyde–alcohol dehydrogenase (AdhE) converts acetyl-CoA to ethanol and is a key enzyme in bacterial alcoholic fermentation. AdhE forms spirosomes and, here, the authors present the cryoEM structures of compact and extended E.coli AdhE spirosomes and show that the extended conformation is the catalytically active form of the enzyme and discuss mechanistic implications.

    • Pauline Pony
    • , Chiara Rapisarda
    •  & Rémi Fronzes
  • Article
    | Open Access

    Lipin/Pah phosphatidic acid phosphatases generate diacylglycerol to regulate triglyceride synthesis and cellular signaling. Here authors determine structures of Tetrahymena thermophila Pah2 and identify an N-terminal amphipathic helix essential for membrane association.

    • Valerie I. Khayyo
    • , Reece M. Hoffmann
    •  & Michael V. Airola
  • Article
    | Open Access

    N-terminal glycine myristoyl transferases (NMTs) catalyse the myristoylation of eukaryotic proteins. Here, the authors provide insights into the catalytic mechanism of NMTs by determining the crystal structures of human NMT1 in complex with reactive cognate lipid and peptide substrates and further show that NMT1 also catalyses the acylation of N-terminal lysines.

    • Cyril Dian
    • , Inmaculada Pérez-Dorado
    •  & Carmela Giglione
  • Article
    | Open Access

    Metabolic engineering is often hampered by non-linear kinetics and allosteric regulatory mechanisms. Here, the authors construct a quantitative model for the pentose degradation Weimberg pathway in Caulobacter crescentus and demonstrate its biotechnological applications in cell-free system and standard metabolic engineering.

    • Lu Shen
    • , Martha Kohlhaas
    •  & Bettina Siebers
  • Article
    | Open Access

    Lysine fatty acylation is an important protein posttranslational modification but mammalian lysine fatty acyl transferases have remained unknown so far. Here the authors report that the human N-terminal glycine myristoyltransferases 1 and 2 catalyze the addition of myristoyl chains to specific lysine residues and show that they myristoylate ARF6 lysine 3, which explains the unusual membrane binding properties of ARF6.

    • Tatsiana Kosciuk
    • , Ian R. Price
    •  & Hening Lin
  • Article
    | Open Access

    Glucuronoyl esterases have the potential to be used in the biocatalytic conversion of lignin-carbohydrate complexes to obtain pure lignin for downstream biofuel conversion. Here the authors present a detailed structural analysis of the glucuronoyl esterase from Cerrena unicolor, providing the basis for its activity on natural substrate and for how lignin can be selectively separated from lignocellulosic materials.

    • Heidi A. Ernst
    • , Caroline Mosbech
    •  & Sine Larsen
  • Article
    | Open Access

    Production of inflammatory mediators by M1-polarized macrophages is thought to rely on suppression of mitochondrial metabolism in favor of glycolysis. Refining this concept, here the authors define metabolic targets of nitric oxide as responsible for the mitochondrial rewiring resulting from polarization.

    • Erika M. Palmieri
    • , Marieli Gonzalez-Cotto
    •  & Daniel W. McVicar
  • Article
    | Open Access

    Respiratory syncytial virus (RSV) is a pathogenic non-segmented negative-sense RNA virus and active RSV polymerase is composed of a 250 kDa large (L) protein and tetrameric phosphoprotein (P). Here, the authors present the 3.67 Å cryo-EM structure of the RSV polymerase (L:P) complex.

    • Dongdong Cao
    • , Yunrong Gao
    •  & Bo Liang
  • Article
    | Open Access

    Understanding enzyme active sites can elucidate fundamental enzymatic reaction pathways and inform designs for synthetic catalysts. Here, authors employ operando X-ray absorption spectroelectrochemistry to assess copper ions in bilirubin oxidase during oxygen reduction electrocatalysis.

    • Lucyano J. A. Macedo
    • , Ayaz Hassan
    •  & Frank N. Crespilho
  • Article
    | Open Access

    The enzyme LspA from the human pathogen Staphylococcus aureus (MRSA) contributes to the integrity and function of the bacterial cell envelope. Here, authors provide crystal structures of LspA in complex with two natural antibiotics, which have profoundly different structures but inhibit LspA in an identical way.

    • Samir Olatunji
    • , Xiaoxiao Yu
    •  & Martin Caffrey
  • Article
    | Open Access

    A robust platform to study modular polyketide synthases (PKSs) in vitro is still unavailable. Here, the authors report the reconstitution of the venemycin PKS, engineer hybrid venemycin/pikromycin PKSs, and obtain much improved yields through employing the updated module boundaries.

    • Takeshi Miyazawa
    • , Melissa Hirsch
    •  & Adrian T. Keatinge-Clay
  • Article
    | Open Access

    Activation of APC/C depends on transient interactions with an activator subunit. Here, the authors show that cellular polyanions trigger APC/C-activator dissociation and that high-affinity substrates block this effect, suggesting that substrate availability controls APC/C-activator binding.

    • Arda Mizrak
    •  & David O. Morgan
  • Article
    | Open Access

    PRDM9 is a PR domain containing histone methyl transferase which expression is normally restricted to the germline that has also been linked to a number of somatic cancers. Here the authors describe the identification of a small molecule that selectivity inhibits the methyltransferase activity of PRDM9 in biochemical and cellular assays

    • Abdellah Allali-Hassani
    • , Magdalena M. Szewczyk
    •  & Masoud Vedadi
  • Article
    | Open Access

    The poly(ADP-ribose) polymerases play a key role in maintaining genomic integrity by detecting DNA damage and mediating repair. Here the authors characterize the kinetics of PARP1 binding to a variety of nucleosomes harbouring DNA double-strand breaks.

    • Deepti Sharma
    • , Louis De Falco
    •  & Curt A. Davey
  • Article
    | Open Access

    The chromosomal kinase JIL-1 is responsible for interphase histone H3S10 phosphorylation and has been proposed to protect active chromatin from heterochromatinisation. Here, the authors show that JIL-1 is stabilized and anchored to active genes and telomeric transposons by JASPer, which binds to H3K36me3 nucleosomes via its PWWP domain.

    • Christian Albig
    • , Chao Wang
    •  & Catherine Regnard