Search

Cobalamin (Cbl)-dependent radical S-adenosyl-L-methionine (SAM) proteins constitute the largest collection of the radical SAM superfamily that has hundreds of thousands of individual members. Many of these proteins are involved in the biosynthesis of pharmaceutically important natural products to catalyze chemically demanding reactions. In the biosynthetic pathway of chuangxinmycin (CXM), a unique indole alkaloid antibiotic with potent anti-infective activity, functionalization of the characteristic thiopyrano[4,3,2-cd]indole scaffold by regio- and stereoselective C3-methylation is believed to rely on a Cbl-dependent radical process, which, however, remained to be reconstituted biochemically. We here report the dissection of this enzymatic process, which requires the incorporation of Cxm8, a Cbl-dependent radical SAM protein, with Cxm9, a DUF5825 family protein that shares no homology to any proteins of known functions. Cxm8 and Cxm9 function together by forming an unexpected heterodimeric complex that selectively catalyzes C3-methylation of the tricyclic indole-S-hetero ring system in a successive manner, achieving CXM and a recently identified, C3-dimethylated congener. Detailed biochemical characterization, isotope labeling, structural simulation and bioinformatics analysis rationalized the catalysis of the Cxm8/Cxm9 complex and particularly the necessity of the DUF5825 protein for C3-methylase activity. This is the first example that a Cbl-dependent protein acts with a partner to exhibit radical SAM activity.

The X-ray spectral state transitions of Galactic black hole transients (GBHTs) are often linked to the changes in the mass accretion rate. A narrow distribution of transition luminosity in terms of the Eddington ratio has been found in previous studies of GBHTs based on RXTE data (Maccarone, 2003, A&A, 409, 697; Vahdat Motlagh et al., 2019, MNRAS, 485, 2744) and this Eddington ratio at the transition is often used in recent studies with instruments such as Swift/XRT and NICER/XTI, covering soft energies below 1 to 10 keV. However, the X-ray states characterized by spectral parameters may have different definitions depending on the energy ranges adopted in the spectral analysis, leaving the question of whether the distribution of transition luminosity obtained with RXTE remains the same when we use the instruments covering softer energy bands. In this work, we investigated the X-ray state evolutions and the variations of luminosities of eight outbursts of seven GBHTs. We found that the bolometric luminosity of the power-law component was tightly constrained to $\sim\! 1.3\%$Eddington luminosity at index transition when the photon index starts to decrease towards the hard state, which is consistent with the previous RXTE results (Vahdat Motlagh et al. 2019, MNRAS, 485, 2744; Kalemci et al. 2013, ApJ, 779, 95). Moreover, the tightest clustering was found to be the power-law luminosity right after the start of disk recession, with a mean logarithmic Eddington ratio of −1.84 ± 0.28. In addition, our results suggest that the disk recession starts after the bolometric disk luminosity drops below 1% Eddington luminosity.

We developed a high-speed image reduction pipeline using Graphics Processing Units (GPUs) as hardware accelerators. Astronomers desire to detect the emission measure counterpart of gravitational-wave sources as soon as possible and to share in the systematic follow-up observation. Therefore, high-speed image processing is important. We developed a new image-reduction pipeline for our robotic telescope system, which uses a GPU via the Python package CuPy for high-speed image processing. As a result, the new pipeline has increased in processing speed by more than 40 times compared with the current one, while maintaining the same functions.

Mitomycins make up a class of natural molecules produced by Streptomyceswith strong antibacterial and antitumor activities. MitM is a key postmitosane modification enzyme involved in mitomycin biosynthesis in Streptomyces caespitosus. This protein was previously suggested to catalyze the aziridinium methylation of mitomycin A and the mitomycin intermediate 9a-demethyl-mitomycin A as an N-methyltransferase. The structural basis for MitM to recognize cofactor S-adenosyl-l-methionine (SAM) and substrate mitomycin A is unknown. Here, we determined the crystal structures of apo-MitM and MitM-mitomycin A-S-adenosylhomocysteine (SAH) ternary complexes with resolutions of 2.23 and 2.80 Å, respectively. We found that MitM adopts a class I SAM-dependent methyltransferase fold and forms a homodimer in solution. Conformational changes in a series of residues involved in the formation of active pockets assist MitM in binding SAH and mitomycin A. In particular, the 28ALGAASLGE36loop changes most significantly. When mitomycin A binds, the bending direction of this loop is reversed, changing the entrance of the active site from open to closed. This study provides structural insights into MitM’s involvement in the postmitosane stage of mitomycin biosynthesis and provides a template for the engineering of methyltransferases.

Mitomycins are a family of naturally occurring, potent alkylating agents in which the C member has been clinically used for cancer chemotherapy for over 5 decades. In Streptomyces caespitosus, mitomycins are derived from an N-glycoside composed of a 3-amino-5-hydroxybenzoic acid (AHBA) unit and a d-glucosamine (GlcN) unit; however, how this N-glycoside is formed and rearranged to a mitosane, for example, the compact polycyclic ring system of mitomycin C, remains elusive. Benefiting from the development of a method used to trace the mitomycin intermediates that accumulate on an acyl carrier protein (ACP), we here dissect the enzymatic steps for AHBA–GlcN formation and processing to underlie the mitosane structure. Following the N-glycosylation of AHBA with activated N-acetyl-GlcN, deacetylation occurs on ACP to provide AHBA–GlcN. Then, the sugar portion of this N-glycoside is transformed into a linear aminodiol that terminates with an epoxyethane, yielding an ACP-channeled intermediate that is ready for mitosane formation through crosslinking between the AHBA and linearized sugar units. This transformation is unusual and relies on the functional association of a dihydronicotinamide adenine dinucleotide (phosphate)-dependent protein with a radical S-adenosyl-l-methionine protein. Characterization of these ACP-based enzymatic steps for AHBA–GlcN formation and processing sheds light on the poorly understood biosynthetic pathway of mitomycins.

J Li, L Bao, ZJ Xia and KY Ding contributed equally to this study.

Jing Li, Li Bao and Zhong-jun Xia contributed equally to this study.

No relevant conflicts of interest to declare.Ixazomib is an oral proteasome inhibitor that approved for the use in patients with relapsed/refractory multiple myeloma (RRMM). Here in this abstract, I will present data on real-life practice of the use of ixazomib in newly diagnosed multiple myeloma.

No relevant conflicts of interest to declare.

We report on the X-ray spectral analysis and time evolution of GRS 1739−278 during its 2014 outburst, based on MAXI/GSC and Swift/XRT observations. Over the course of the outburst, a transition from the low/hard state to the high/soft state and then back to the low/hard state was seen. During the high/soft state, the innermost disk temperature mildly decreased, while the innermost radius estimated with the multi-color disk model remained constant at ∼18 (D/8.5 kpc)(cos i/cos 30°)−1/2km, where Dis the source distance and iis the inclination of observation. This small innermost radius of the accretion disk suggests that the central object is more likely to be a Kerr black hole rather than a Schwardzschild black hole. Applying a relativistic disk emission model to the high/soft state spectra, a mass upper limit of 18.3 M⊙was obtained based on the inclination limit i< 60° for an assumed distance of 8.5 kpc. Using the empirical relation of the transition luminosity to the Eddington limit, the mass is constrained to 4.0–18.3 M⊙for the same distance. The mass can be further constrained to be no larger than 9.5 M⊙by adopting the constraints based on the fits to the NuSTAR spectra with relativistically blurred disk reflection models (Miller et al. 2015, ApJ, 799, L6).