A5057528779- Peptidase Inhibition and Analysis
- Plant biochemistry and biosynthesis
- Cancer Research and Treatments
- Neuroendocrine Tumor Research Advances
- Microbial Natural Products and Biosynthesis
- Adenosine and Purinergic Signaling
- Antioxidant Activity and Oxidative Stress
- Ubiquitin and proteasome pathways
- Microbial Metabolic Engineering and Bioproduction
- Enzyme Structure and Function
- Photosynthetic Processes and Mechanisms
- Natural product bioactivities and synthesis
- Biochemical and Molecular Research
- Multiple Myeloma Research and Treatments
- Pharmacological Effects of Natural Compounds
- Cancer-related Molecular Pathways
- Neuropeptides and Animal Physiology
- Pancreatic and Hepatic Oncology Research
- Protein Degradation and Inhibitors
- Computational Drug Discovery Methods
- Advanced Breast Cancer Therapies
- Crystallization and Solubility Studies
- Lung Cancer Research Studies
- Wnt/β-catenin signaling in development and cancer
- X-ray Diffraction in Crystallography
Merck (Germany)
2011-2023
Ono Pharmaceutical (United States)
2021
Research & Development Institute
2021
Merck Serono (Italy)
2013
Technical University of Munich
2003-2012
Center for Integrated Protein Science Munich
2008-2010
Leibniz-Institute for Food Systems Biology at the Technical University of Munich
2006
Universität Hamburg
2006
ETH Zurich
2002-2004
Freie Universität Berlin
2004
Isopentenyl diphosphate and dimethylallyl serve as the universal precursors for biosynthesis of terpenes. Although their by means mevalonate has been studied in detail, a second biosynthetic pathway formation 1-deoxy- d -xylulose 5-phosphate discovered only recently plants certain eubacteria. Earlier vivo experiments with recombinant Escherichia coli strains showed that exogenous can be converted into 1-hydroxy-2-methyl-2-( E )-butenyl 4-diphosphate consecutive action enzymes specified xylB...
2 -C -methylerythritol 4-phosphate has been established recently as an intermediate of the deoxyxylulose phosphate pathway used for biosynthesis terpenoids in plants and many microorganisms. We show that enzyme isolated from cell extract Escherichia coli converts into 4-diphosphocytidyl-2 by reaction with CTP. The is specified hitherto unannotated ORF ygbP E. . cognate protein was obtained pure form a recombinant hyperexpression strain harboring plasmid gene under control T5 promoter lac...
In many microorganisms, the putative orthologs of Escherichia coli ygbB gene are tightly linked or fused to ygbP , which has been shown earlier be involved in terpenoid biosynthesis. The E. was expressed a recombinant strain and direct synthesis soluble, 17-kDa polypeptide. protein found convert 4-diphosphocytidyl-2C-methyl- d -erythritol 2-phosphate into 2C-methyl- 2,4-cyclodiphosphate CMP. structure reaction product established by NMR spectroscopy using 13 C-labeled substrate samples....
Earlier in vivo studies have shown that the sequential action of IspG and IspH proteins is essential for reductive transformation 2C-methyl-d-erythritol 2,4-cyclodiphosphate into dimethylallyl diphosphate isopentenyl via 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate. A recombinant fusion protein comprising maltose binding domains was purified from a Escherichia coli strain. The failed to transform 4-diphosphate, but catalytic activity could be restored by addition crude cell extract an...
A comparative analysis of all published complete genomes indicated that the putative orthologs unannotated ychB gene Escherichia coli follow distribution dxs , dxr and ygbP genes, which have been shown to specify enzymes deoxyxylulose phosphate pathway terpenoid biosynthesis, thus suggesting hypothetical YchB protein also is involved in pathway. To test this hypothesis, E. was expressed a homologous host. The recombinant purified homogeneity phosphorylate 4-diphosphocytidyl-2C-methyl- d...
Recombinant Escherichia coli cells engineered for the expression of xylB gene in conjunction with genes nonmevalonate pathway were supplied 13 C-labeled 1-deoxy- d -xylulose. Cell extracts analyzed directly by NMR spectroscopy. 2 C -methyl- -erythritol 2,4-cyclodiphosphate was detected at high levels expressing , ispC ispD ispE, and ispF . The additional gcpE afforded 1-hydroxy-2-methyl-2-( E )-butenyl 4-diphosphate as an intermediate pathway. Hypothetical mechanisms involving conserved...
The Mediator complex-associated cyclin-dependent kinase CDK8 has been implicated in human disease, particularly colorectal cancer where it reported as a putative oncogene. Here we report the discovery of 109 (CCT251921), potent, selective, and orally bioavailable inhibitor with equipotent affinity for CDK19. We describe structure-based design approach leading to 3,4,5-trisubstituted-2-aminopyridine series present application physicochemical property analyses successfully reduce vivo...
The mediator complex-associated cyclin dependent kinase CDK8 regulates β-catenin-dependent transcription following activation of WNT signaling. Multiple lines evidence suggest may act as an oncogene in the development colorectal cancer. Here we describe successful optimization imidazo-thiadiazole series inhibitors that was identified a high-throughput screening campaign and further progressed by structure-based design. In several cycles, improved microsomal stability, potency, selectivity....
Mediator-associated kinases CDK8/19 are context-dependent drivers or suppressors of tumorigenesis. Their inhibition is predicted to have pleiotropic effects, but it unclear whether this will impact on the clinical utility inhibitors. We discovered two series potent chemical probes with high selectivity for CDK8/19. Despite pharmacodynamic evidence robust on-target activity, compounds exhibited modest, though significant, efficacy against human tumor lines and patient-derived xenografts....
2-C-Methyl-d-erythritol 4-phosphate synthase (IspC) is the first enzyme committed to isoprenoid biosynthesis in methylerythritol phosphate pathway, which represents an alternative route classical mevalonate pathway. As it present many pathogens and plants, but not man, this pathway has attracted considerable interest as a target for novel antibiotics herbicides. Fosmidomycin specific high-affinity inhibitor of IspC. Very recently, its anti-malaria activity man been demonstrated clinical...
Earlier in vivo studies showed the involvement of IspH protein conversion 1-hydroxy-2-methyl-2-( E )-butenyl 4-diphosphate into isopentenyl diphosphate (IPP) and dimethylallyl (DMAPP). We have demonstrated now that cell extract an Escherichia coli strain engineered for hyperexpression ispH ( lytB ) gene catalyzes vitro IPP DMAPP. The reaction requires NADH, FAD, divalent cations (preferably Co 2+ ), probably one or more as-yet-unidentified proteins. low intrinsic catalytic activities...
Isopentenyl diphosphate (IPP) and dimethylallyl (DMAPP) represent the two central intermediates in biosynthesis of isoprenoids. The recently discovered deoxyxylulose 5-phosphate pathway generates a mixture IPP DMAPP its final step by reductive dehydroxylation 1-hydroxy-2-methyl-2-butenyl 4-diphosphate. This conversion is catalyzed IspH protein comprising iron-sulfur cluster as electron transfer cofactor active site. five crystal structures complex with substrate, converted products PP i...
WNT signaling is frequently deregulated in malignancy, particularly colon cancer, and plays a key role the generation maintenance of cancer stem cells. We report discovery optimization 3,4,5-trisubstituted pyridine 9 using high-throughput cell-based reporter assay pathway activity. demonstrate twisted conformation about pyridine–piperidine bond by small-molecule X-ray crystallography. Medicinal chemistry to maintain this conformation, cognisant physicochemical properties likely good cell...
A hypothetical gene with similarity to the ispD of Escherichia coli was cloned from Arabidopsis thaliana cDNA. The ORF 909 bp specifies a protein 302 amino acid residues. cognate chromosomal consists 2,071 and comprises 11 introns size range 78-202 bp. fragment comprising residues 76-302 expressed in recombinant E. strain. purified homogeneity shown catalyze formation 4-diphosphocytidyl-2C-methyl-d-erythritol 2C-methyl-d-erythritol 4-phosphate specific activity 67 micromol small middle...
The ispH gene of Escherichia coli specifies an enzyme catalyzing the conversion 1-hydroxy-2-methyl-2-(E)-butenyl diphosphate into a mixture isopentenyl (IPP) and dimethylallyl (DMAPP) in nonmevalonate isoprenoid biosynthesis pathway. implementation cassette directing overexpression isc operon involved assembly iron−sulfur clusters strain engineered for expression increased catalytic activity IspH protein anaerobically purified from this by factor at least 200. For maximum activity,...
The putative catalytic domain (residues 81–401) of a predicted tomato protein with similarity to 4-diphosphocytidyl-2- C -methyl- d -erythritol kinase Escherichia coli was expressed in recombinant E. strain. purified homogeneity and shown catalyze the phosphorylation position 2 hydroxy group at rate 33 μmol⋅mg −1 ⋅min . structure reaction product, 2-phosphate, established by NMR spectroscopy. Divalent metal ions, preferably Mg 2+ , are required for activity. Neither enzyme nor ortholog...
The terminal step of the non-mevalonate pathway terpene biosynthesis is catalyzed by IspH (see scheme). In crystal structure from E. coli, a bound inorganic diphosphate ligand occupies position residue substrate. Together with mutation studies and theoretical calculations, these data support mechanism which analogous to Birch reduction allylic alcohols. Detailed facts importance specialist readers are published as ”Supporting Information”. Such documents peer-reviewed, but not copy-edited or...