A DECam Search for Explosive Optical Transients Associated with IceCube Neutrino Alerts

In this work, we investigate the likelihood of association between real-time, neutrino alerts with teraelectronvolt to petaelectronvolt energy from IceCube and optical counterparts in the form of core-collapse supernovae (CC SNe). The optical follow-up of IceCube alerts requires two main instrumental capabilities: (1) deep imaging, since 73% of neutrinos would come from CC SNe at redshifts z > 0.3, and (2) a large field of view (FoV), since typical IceCube muon neutrino pointing accuracy is on the order of ~1 deg. With Blanco/DECam (gri to 24th magnitude and 2.2 deg diameter FoV), we performed a triggered optical follow-up observation of two IceCube alerts, IC170922A and IC171106A, on six nights during the three weeks following each alert. For the IC170922A (IC171106A) follow-up observations, we expect that 12.1% (9.5%) of coincident CC SNe at z lesssim 0.3 are detectable, and that, on average, 0.23 (0.07) unassociated SNe in the neutrino 90% containment regions also pass our selection criteria. We find two candidate CC SNe that are temporally coincident with the neutrino alerts in the FoV, but none in the 90% containment regions, a result that is statistically consistent with expected rates of background CC SNe for these observations. If CC SNe are the dominant source of teraelectronvolt to petaelectronvolt neutrinos, we would expect an excess of coincident CC SNe to be detectable at the 3σ confidence level using DECam observations similar to those of this work for ~60 (~200) neutrino alerts with (without) redshift information for all candidates

Structure and Function of NzeB, a Versatile C–C and C–N Bond-Forming Diketopiperazine Dimerase

The dimeric diketopiperazine (DKPs) alkaloids are a diverse family of natural products (NPs) whose unique structural architectures and biological activities have inspired the development of new synthetic methodologies to access these molecules. However, catalyst-controlled methods that enable the selective formation of constitutional and stereoisomeric dimers from a single monomer are lacking. To resolve this long-standing synthetic challenge, we sought to characterize the biosynthetic enzymes that assemble these NPs for application in biocatalytic syntheses. Genome mining enabled identification of the cytochrome P450, NzeB (Streptomyces sp. NRRL F-5053), which catalyzes both intermolecular carbon-carbon (C-C) and carbon-nitrogen (C-N) bond formation. To identify the molecular basis for the flexible site-selectivity, stereoselectivity, and chemoselectivity of NzeB, we obtained high-resolution crystal structures (1.5 Å) of the protein in complex with native and non-native substrates. This, to our knowledge, represents the first crystal structure of an oxidase catalyzing direct, intermolecular C-H amination. Site-directed mutagenesis was utilized to assess the role individual active-site residues play in guiding selective DKP dimerization. Finally, computational approaches were employed to evaluate plausible mechanisms regarding NzeB function and its ability to catalyze both C-C and C-N bond formation. These results provide a structural and computational rationale for the catalytic versatility of NzeB, as well as new insights into variables that control selectivity of CYP450 diketopiperazine dimerases

Structure and Function of NzeB, a Versatile C–C and C–N Bond-Forming Diketopiperazine Dimerase

© 2020 American Chemical Society. The dimeric diketopiperazine (DKPs) alkaloids are a diverse family of natural products (NPs) whose unique structural architectures and biological activities have inspired the development of new synthetic methodologies to access these molecules. However, catalyst-controlled methods that enable the selective formation of constitutional and stereoisomeric dimers from a single monomer are lacking. To resolve this long-standing synthetic challenge, we sought to characterize the biosynthetic enzymes that assemble these NPs for application in biocatalytic syntheses. Genome mining enabled identification of the cytochrome P450, NzeB (Streptomyces sp. NRRL F-5053), which catalyzes both intermolecular carbon-carbon (C-C) and carbon-nitrogen (C-N) bond formation. To identify the molecular basis for the flexible site-selectivity, stereoselectivity, and chemoselectivity of NzeB, we obtained high-resolution crystal structures (1.5 Å) of the protein in complex with native and non-native substrates. This, to our knowledge, represents the first crystal structure of an oxidase catalyzing direct, intermolecular C-H amination. Site-directed mutagenesis was utilized to assess the role individual active-site residues play in guiding selective DKP dimerization. Finally, computational approaches were employed to evaluate plausible mechanisms regarding NzeB function and its ability to catalyze both C-C and C-N bond formation. These results provide a structural and computational rationale for the catalytic versatility of NzeB, as well as new insights into variables that control selectivity of CYP450 diketopiperazine dimerases

Fungal indole alkaloid biogenesis through evolution of a bifunctional reductase/Diels–Alderase

Prenylated indole alkaloids such as the calmodulin-inhibitory malbrancheamides and anthelmintic paraherquamides possess great structural diversity and pharmaceutical utility. Here, we report complete elucidation of the malbrancheamide biosynthetic pathway accomplished through complementary approaches. These include a biomimetic total synthesis to access the natural alkaloid and biosynthetic intermediates in racemic form and in vitro enzymatic reconstitution to provide access to the natural antipode (+)-malbrancheamide. Reductive cleavage of an L-Pro-L-Trp dipeptide from the MalG non-ribosomal peptide synthetase (NRPS) followed by reverse prenylation and a cascade of post-NRPS reactions culminates in an intramolecular [4+2] hetero-Diels-Alder (IMDA) cyclization to furnish the bicyclo[2.2.2]diazaoctane scaffold. Enzymatic assembly of optically pure (+)-premalbrancheamide involves an unexpected zwitterionic intermediate where MalC catalyses enantioselective cycloaddition as a bifunctional NADPH-dependent reductase/Diels-Alderase. The crystal structures of substrate and product complexes together with site-directed mutagenesis and molecular dynamics simulations demonstrate how MalC and PhqE (its homologue from the paraherquamide pathway) catalyse diastereo- and enantioselective cyclization in the construction of this important class of secondary metabolites

Upper Urinary Tract Tumors

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