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Journal of Natural Medicines Jan 2020A new prenylated benzophenone, hypatulin C (1), was isolated from the leaves of Hypericum patulum together with a biogenetically related analog, hypelodin B (2)....
A new prenylated benzophenone, hypatulin C (1), was isolated from the leaves of Hypericum patulum together with a biogenetically related analog, hypelodin B (2). Hypatulin C (1) had a tricyclic [4.3.1.0]-decane moiety substituted by four prenyl groups. The structure of 1 was elucidated by detailed spectroscopic analyses, while the absolute stereochemistries of 1 and 2 were assigned by comparison of their ECD spectra with TDDFT calculated spectra.
Topics: Benzophenones; Hypericum; Molecular Structure; Phloroglucinol; Plant Leaves; Prenylation; Stereoisomerism
PubMed: 31377922
DOI: 10.1007/s11418-019-01350-3 -
Small GTPases Jul 2020Mutant RAS isoforms are the most common oncogenes affecting human cancers. After decades of effort in developing drugs targeting oncogenic RAS-driven cancers, we are... (Review)
Review
Mutant RAS isoforms are the most common oncogenes affecting human cancers. After decades of effort in developing drugs targeting oncogenic RAS-driven cancers, we are still charting an unclear path. Despite recent developments exemplified by KRAS (G12C) inhibitors, direct targeting of mutant RAS remains a difficult endeavor. Inhibiting RAS function by targeting its post-translational prenylation processing has remained an important approach, especially with recent progress on the study of isoprenylcysteine carboxylmethyltransferase (ICMT), the unique enzyme for the last step of prenylation processing of RAS isoforms and other substrates. Inhibition of ICMT has shown efficacy both and in RAS-mutant cancer models. We will discuss the roles of RAS family of proteins in human cancers and the impact of post-prenylation carboxylmethylation on RAS driven tumorigenesis. In addition, we will review what is known of the molecular and cellular impact of ICMT inhibition on cancer cells that underlie its anti-proliferative and pro-apoptosis efficacy.
Topics: Animals; Humans; Methylation; Protein Prenylation; ras Proteins
PubMed: 29261009
DOI: 10.1080/21541248.2017.1415637 -
Applied and Environmental Microbiology May 2021Cyclodipeptide synthases (CDPSs) catalyze the formation of cyclodipeptides using aminoacylated tRNAs as the substrates and have great potential in the production of...
Cyclodipeptide synthases (CDPSs) catalyze the formation of cyclodipeptides using aminoacylated tRNAs as the substrates and have great potential in the production of diverse 2,5-diketopiperazines (2,5-DKPs). Genome mining of NRRL B-24963 revealed a two-gene locus, , encoding CDPS SazA and a unique fused enzyme (SazB) harboring two domains: phytoene synthase-like prenyltransferase (PT) and methyltransferase (MT). Heterologous expression of the gene(s) in J1074 led to the production of four prenylated indole alkaloids, among which streptoazines A to C (compounds 3 to 5) are new compounds. Expression of different gene combinations showed that the SazA catalyzes the formation of (l-Trp-l-Trp) (cWW; compound 1), followed by consecutive prenylation and methylation by SazB. Biochemical assays demonstrated that SazB is a bifunctional enzyme, catalyzing sequential C-3/C-3' prenylation(s) by SazB-PT and N-1/N-1' methylation(s) by SazB-MT. Of note, the substrate selectivity of SazB-PT and SazB-MT was probed, revealing the stringent specificity of SazB-PT but relative flexibility of SazB-MT. Natural products with a 2,5-diketopiperazine (2,5-DKP) skeleton have long sparked interest in drug discovery and development. Recent advances in microbial genome sequencing have revealed that the potential of cyclodipeptide synthase (CDPS)-dependent pathways encoding new 2,5-DKPs are underexplored. In this study, we report the genome mining of a new CDPS-encoding two-gene operon and activation of this cryptic gene cluster through heterologous expression, leading to the discovery of four indole 2,5-DKP alkaloids. The (l-Trp-l-Trp) (cWW)-synthesizing CDPS SazA and the unusual prenyltransferase (PT)-methyltransferase (MT) fused enzyme SazB were characterized. Our results expand the repertoire of CDPSs and associated tailoring enzymes, setting the stage for accessing diverse prenylated alkaloids using synthetic biology strategies.
Topics: Bacterial Proteins; Indole Alkaloids; Metabolic Networks and Pathways; Microorganisms, Genetically-Modified; Peptide Synthases; Prenylation; Streptomyces
PubMed: 33741615
DOI: 10.1128/AEM.02525-20 -
Journal of Agricultural and Food... Feb 2022Prenylated chalcones are found mainly in plants and exhibit diverse biological and pharmacological activities. Some of these compounds are components of food and dietary...
Prenylated chalcones are found mainly in plants and exhibit diverse biological and pharmacological activities. Some of these compounds are components of food and dietary supplements with significant health benefits. In plants, they are derived from chalcones by prenylation with membrane-bound prenyltransferases. In this study, we demonstrate prenylations of 10 chalcones by two fungal prenyltransferases (AtaPT/AnaPT) in the presence of dimethylallyl diphosphate. Eleven mono (- and ) and four diprenylated products (, , , and ) were obtained. Among them, 12 have new structures (, , -, , , , , , , and ). Most of the obtained prenylated chalcones are products of AnaPT and carry prenyl moieties at ring B. Our study provides an excellent example for increasing structural diversity of plant metabolites with microbial enzymes.
Topics: Chalcones; Dimethylallyltranstransferase; Fungi; Plants; Prenylation
PubMed: 35089022
DOI: 10.1021/acs.jafc.1c07786 -
Molecular Neurobiology Dec 2019Early activation of cytosolic NADPH oxidase-2 (Nox2) in diabetes increases retinal ROS production, damaging their mitochondria. The assembly of Nox2 holoenzyme requires...
PURPOSE
Early activation of cytosolic NADPH oxidase-2 (Nox2) in diabetes increases retinal ROS production, damaging their mitochondria. The assembly of Nox2 holoenzyme requires activation of a small molecular weight G protein Rac1. Rac1 activation is regulated by guanine exchange factors and guanine nucleotide-dissociation inhibitors, and post-translational modifications assist in its association with exchange factors and dissociation inhibitors. The goal of this study is to investigate the mechanisms of Rac1 activation in the development of diabetic retinopathy.
METHODS
The levels of the dissociation inhibitor, prenylating enzyme (farnesyltransferase, FNTA), and exchange factor Vav2 were quantified in human retinal endothelial cells, incubated in normal or high glucose for 96 h. The roles of prenylation and Vav2 in Rac1-Nox2-ROS mitochondrial damage were confirmed in FNTA-siRNA-transfected cells and using the Vav2 inhibitor EHop, respectively. Retinal histopathology and functional changes associated with diabetic retinopathy were analyzed in diabetic mice receiving EHop for 6 months. Key parameters of Rac1 activation were confirmed in the retinal microvasculature from human donors with diabetic retinopathy.
RESULTS
In HRECs, glucose increased FNTA and Vav2 and decreased the dissociation inhibitor. FNTA-siRNA and EHop inhibited glucose-induced activation of Rac1-Nox2-ROS signaling. In diabetic mice, EHop ameliorated the development of retinopathy and functional/structural abnormalities and attenuated Rac1-Nox2-mitochondrial damage. Similar alterations in Rac1 regulators were observed in retinal microvasculature from human donors with diabetic retinopathy. In diabetes, Rac1 prenylation and its interactions with Vav2 contribute to Nox2-ROS-mitochondrial damage, and the pharmacological inhibitors to attenuate Rac1 interactions with its regulators could have the potential to halt/inhibit the development of diabetic retinopathy. Graphical Abstract Activation of prenylating enzyme farnesyltransferase (FNTA) in diabetes, prenylates Rac1. The binding of Rac1 with guanine nucleotide-dissociation inhibitor (GDI) is decreased, but its association with the guanine exchange factor, Vav2, is increased, resulting in Rac1 activation. Active Rac1 helps in the assembly of Nox2 holoenzyme, and Nox2 activation increases cytosolic ROS production, damaging the mitochondria. Damaged mitochondria accelerate capillary cell apoptosis, and ultimately, results in the development of diabetic retinopathy.
Topics: Aged; Animals; Diabetic Retinopathy; Endothelial Cells; Glucose; Humans; Mice, Inbred C57BL; Microvessels; Middle Aged; NADPH Oxidase 2; Oxidative Stress; Prenylation; Proto-Oncogene Proteins c-vav; Tissue Donors; rac1 GTP-Binding Protein
PubMed: 31300985
DOI: 10.1007/s12035-019-01696-5 -
Analytical Chemistry Oct 2021Protein prenylation is an important post-translational modification that regulates protein interactions, localizations, and signaling pathways in normal functioning of...
Protein prenylation is an important post-translational modification that regulates protein interactions, localizations, and signaling pathways in normal functioning of eukaryotic cells. It is also a critical step in the oncogenic developments of various cancers. Direct identification of native protein prenylation by mass spectrometry (MS) has been challenging due to high hydrophobicity and the lack of an efficient enrichment technique. Prior MS studies of prenylation revealed that prenyl peptides readily generate high-intensity fragments after neutral loss of the prenyl group (R group), and more recent investigation of oxidized prenyl peptides discovered more consistent neutral loss of the oxidized prenyl group (RSOH group). Here, a dual-stage neutral loss MS (DS-NLMS3)-based strategy is therefore developed by combining both gas-phase cleavable properties of the prenyl thioether bond and mono-oxidized thioether to improve the large-scale identification of prenylation. Both neutral losses can individually and distinctively confirm the prenylation type in MS and the sequence of the prenyl peptide upon targeted MS fragmentation. This dual-faceted NLMS3 strategy significantly improves the confidence in the identification of protein prenylation from large-scale samples, which enables the unambiguous identification of prenylated sites of the spiked low-abundance farnesyl peptide and native prenyl proteins from mouse macrophage cells, even without prior enrichment during sample preparation. The ease of incorporating this strategy into the prenylation study workflow and minimum disruption to the biological lipidome are advantageous for unraveling unknown native protein prenylation and further developments in profiling and quantifying prenylome.
Topics: Animals; Mice; Protein Prenylation
PubMed: 34558911
DOI: 10.1021/acs.analchem.1c01617 -
Biological & Pharmaceutical Bulletin 2024CsPT4 is an aromatic prenyltransferase that synthesizes cannabigerolic acid (CBGA), the key intermediate of cannabinoid biosynthesis in Cannabis sativa, from olivetolic...
CsPT4 is an aromatic prenyltransferase that synthesizes cannabigerolic acid (CBGA), the key intermediate of cannabinoid biosynthesis in Cannabis sativa, from olivetolic acid (OA) and geranyl diphosphate (GPP). CsPT4 has a catalytic potential to produce a variety of CBGA analogs via regioselective C-prenylation of aromatic substrates having resorcylic acid skeletons including bibenzyl 2,4-dihydroxy-6-phenylethylbenzoic acid (DPA). In this study, we further investigated the substrate specificity of CsPT4 using phlorocaprophenone (PCP) and 2',4',6'-trihydroxydihydrochalcone (THDC), the isomers of OA and DPA, respectively, and demonstrated that CsPT4 catalyzed both C-prenylation and O-prenylation reactions on PCP and THDC that share acylphloroglucinol substructures. Interestingly, the kinetic parameters of CsPT4 for these substrates differed depending on whether they underwent C-prenylation or O-prenylation, suggesting that this enzyme utilized different substrate-binding modes suitable for the respective reactions. Aromatic prenyltransferases that catalyze O-prenylation are rare in the plant kingdom, and CsPT4 was notable for altering the reaction specificity between C- and O-prenylations depending on the skeletons of aromatic substrates. We also demonstrated that enzymatically synthesized geranylated acylphloroglucinols had potent antiausterity activity against PANC-1 human pancreatic cancer cells, with 4'-O-geranyl THDC being the most effective. We suggest that CsPT4 is a valuable catalyst to generate biologically active C- and O-prenylated molecules that could be anticancer lead compounds.
Topics: Humans; Cannabis; Dimethylallyltranstransferase; Prenylation; Catalysis; Substrate Specificity
PubMed: 38369346
DOI: 10.1248/bpb.b23-00868 -
Journal of Natural Products Oct 2019Six new prenylated acylphloroglucinols with menthane moieties, hypascyrins A-E (-) and -hyphenrone J (), together with four known analogues, were isolated from roots....
Six new prenylated acylphloroglucinols with menthane moieties, hypascyrins A-E (-) and -hyphenrone J (), together with four known analogues, were isolated from roots. Detailed spectroscopic data analyses resulted in the assignment of their structures. The absolute configuration of was deduced by experimental and calculated ECD data, while those of - were assigned by ECD data analyses as well as chemical conversions. Hypascyrins A (), C (), and E () and -hyphenrone J () exhibited antimicrobial activity against MRSA (MIC values of 4.0, 8.0, 2.0, and 4.0 μM, respectively) and (MIC values of 4.0, 4.0, 2.0, and 4.0 μM, respectively).
Topics: Anti-Infective Agents; Bacillus subtilis; Hypericum; Methicillin-Resistant Staphylococcus aureus; Phenols; Phloroglucinol; Plant Roots; Prenylation
PubMed: 31596079
DOI: 10.1021/acs.jnatprod.9b00354 -
Journal of Natural Products Mar 2024Prenyltransferases (PTs) are involved in the biosynthesis of a multitude of pharmaceutically and agriculturally important plant, bacterial, and fungal compounds....
Prenyltransferases (PTs) are involved in the biosynthesis of a multitude of pharmaceutically and agriculturally important plant, bacterial, and fungal compounds. Although numerous prenylated compounds have been isolated from Basidiomycota (mushroom-forming fungi), knowledge of the PTs catalyzing the transfer reactions in this group of fungi is scarce. Here, we report the biochemical characterization of an - and -prenylating dimethylallyltryptophan synthase (DMATS)-like enzyme LpTyrPT from the scurfy deceiver This PT transfers dimethylallyl moieties to l-tyrosine at the - position and to l-tryptophan at atom C-7 and represents the first basidiomycete l-tyrosine PT described so far. Phylogenetic analysis of PTs in fungi revealed that basidiomycete l-tyrosine PTs have evolved independently from their ascomycete counterparts and might represent the evolutionary origin of PTs acting on phenolic compounds in secondary metabolism.
Topics: Dimethylallyltranstransferase; Phylogeny; Tyrosine; Basidiomycota; Prenylation
PubMed: 38231181
DOI: 10.1021/acs.jnatprod.3c01120 -
Bioorganic Chemistry Feb 2016Protein prenylation is a posttranslational modification catalyzed by prenyltransferases involving the attachment of farnesyl or geranylgeranyl groups to residues near... (Review)
Review
Protein prenylation is a posttranslational modification catalyzed by prenyltransferases involving the attachment of farnesyl or geranylgeranyl groups to residues near the C-termini of proteins. This irreversible covalent modification is important for membrane localization and proper signal transduction. Here, the use of isoprenoid analogues for studying prenylated proteins is reviewed. First, experiments with analogues containing small fluorophores that are alternative substrates for prenyltransferases are described. Those analogues have been useful for quantifying binding affinity and for the production of fluorescently labeled proteins. Next, the use of analogues that incorporate biotin, bioorthogonal groups or antigenic moieties is described. Such probes have been particularly useful for identifying proteins that are naturally prenylated within mammalian cells. Overall, the use of isoprenoid analogues has contributed significantly to the understanding of protein prenlation.
Topics: Alkyl and Aryl Transferases; Fluorescent Dyes; Molecular Probes; Protein Prenylation; Proteins; Proteomics; Terpenes
PubMed: 26709869
DOI: 10.1016/j.bioorg.2015.12.003