scholarly journals Catabolic Reductive Dehalogenase Substrate Complex Structures Underpin Rational Repurposing of Substrate Scope

2020 ◽  
Vol 8 (9) ◽  
pp. 1344
Author(s):  
Tom Halliwell ◽  
Karl Fisher ◽  
Karl A. P. Payne ◽  
Stephen E. J. Rigby ◽  
David Leys

Reductive dehalogenases are responsible for the reductive cleavage of carbon-halogen bonds during organohalide respiration. A variety of mechanisms have been proposed for these cobalamin and [4Fe-4S] containing enzymes, including organocobalt, radical, or cobalt-halide adduct based catalysis. The latter was proposed for the oxygen-tolerant Nitratireductor pacificus pht-3B catabolic reductive dehalogenase (NpRdhA). Here, we present the first substrate bound NpRdhA crystal structures, confirming a direct cobalt–halogen interaction is established and providing a rationale for substrate preference. Product formation is observed in crystallo due to X-ray photoreduction. Protein engineering enables rational alteration of substrate preference, providing a future blue print for the application of this and related enzymes in bioremediation.

Author(s):  
V. Serin ◽  
K. Hssein ◽  
G. Zanchi ◽  
J. Sévely

The present developments of electron energy analysis in the microscopes by E.E.L.S. allow an accurate recording of the spectra and of their different complex structures associated with the inner shell electron excitation by the incident electrons (1). Among these structures, the Extended Energy Loss Fine Structures (EXELFS) are of particular interest. They are equivalent to the well known EXAFS oscillations in X-ray absorption spectroscopy. Due to the EELS characteristic, the Fourier analysis of EXELFS oscillations appears as a promising technique for the characterization of composite materials, the major constituents of which are low Z elements. Using EXELFS, we have developed a microstructural study of carbon fibers. This analysis concerns the carbon K edge, which appears in the spectra at 285 eV. The purpose of the paper is to compare the local short range order, determined by this way in the case of Courtauld HTS and P100 ex-polyacrylonitrile carbon fibers, which are high tensile strength (HTS) and high modulus (HM) fibers respectively.


2021 ◽  
Vol 49 (5) ◽  
pp. 2684-2699
Author(s):  
Ka-Weng Ieong ◽  
Gabriele Indrisiunaite ◽  
Arjun Prabhakar ◽  
Joseph D Puglisi ◽  
Måns Ehrenberg

Abstract We used quench flow to study how N6-methylated adenosines (m6A) affect the accuracy ratio between kcat/Km (i.e. association rate constant (ka) times probability (Pp) of product formation after enzyme-substrate complex formation) for cognate and near-cognate substrate for mRNA reading by tRNAs and peptide release factors 1 and 2 (RFs) during translation with purified Escherichia coli components. We estimated kcat/Km for Glu-tRNAGlu, EF-Tu and GTP forming ternary complex (T3) reading cognate (GAA and Gm6AA) or near-cognate (GAU and Gm6AU) codons. ka decreased 10-fold by m6A introduction in cognate and near-cognate cases alike, while Pp for peptidyl transfer remained unaltered in cognate but increased 10-fold in near-cognate case leading to 10-fold amino acid substitution error increase. We estimated kcat/Km for ester bond hydrolysis of P-site bound peptidyl-tRNA by RF2 reading cognate (UAA and Um6AA) and near-cognate (UAG and Um6AG) stop codons to decrease 6-fold or 3-fold by m6A introduction, respectively. This 6-fold effect on UAA reading was also observed in a single-molecule termination assay. Thus, m6A reduces both sense and stop codon reading accuracy by decreasing cognate significantly more than near-cognate kcat/Km, in contrast to most error inducing agents and mutations, which increase near-cognate at unaltered cognate kcat/Km.


ChemInform ◽  
2010 ◽  
Vol 24 (2) ◽  
pp. no-no
Author(s):  
R. E. MORRIS ◽  
W. T. A. HARRISON ◽  
J. M. NICOL ◽  
A. P. WILKINSON ◽  
A. K. CHEETHAM

2008 ◽  
Vol 17 (7) ◽  
pp. 1175-1187 ◽  
Author(s):  
Valentina Tereshko ◽  
Serdar Uysal ◽  
Akiko Koide ◽  
Katrina Margalef ◽  
Shohei Koide ◽  
...  

Author(s):  
Raj R. V. Dhanaraj ◽  
Jim E. Pitts ◽  
Phil Nugent ◽  
Poonsook Orprayoon ◽  
Jon B. Cooper ◽  
...  

IUCrJ ◽  
2015 ◽  
Vol 2 (5) ◽  
pp. 498-510 ◽  
Author(s):  
Christer B. Aakeröy ◽  
Christine L. Spartz ◽  
Sean Dembowski ◽  
Savannah Dwyre ◽  
John Desper

As halogen bonds gain prevalence in supramolecular synthesis and materials chemistry, it has become necessary to examine more closely how such interactions compete with or complement hydrogen bonds whenever both are present within the same system. As hydrogen and halogen bonds have several fundamental features in common, it is often difficult to predict which will be the primary interaction in a supramolecular system, especially as they have comparable strength and geometric requirements. To address this challenge, a series of molecules containing both hydrogen- and halogen-bond donors were co-crystallized with various monotopic, ditopic symmetric and ditopic asymmetric acceptor molecules. The outcome of each reaction was examined using IR spectroscopy and, whenever possible, single-crystal X-ray diffraction. 24 crystal structures were obtained and subsequently analyzed, and the synthon preferences of the competing hydrogen- and halogen-bond donors were rationalized against a background of calculated molecular electrostatic potential values. It has been shown that readily accessible electrostatic potentials can offer useful practical guidelines for predicting the most likely primary synthons in these co-crystals as long as the potential differences are weighted appropriately.


2005 ◽  
Vol 353 (4) ◽  
pp. 838-846 ◽  
Author(s):  
Mirjam Czjzek ◽  
Alon Ben David ◽  
Tsafrir Bravman ◽  
Gil Shoham ◽  
Bernard Henrissat ◽  
...  

2020 ◽  
Vol 151 (11) ◽  
pp. 1673-1679
Author(s):  
Nataliya L. Gulay ◽  
Yaroslav M. Kalychak ◽  
Maximilian K. Reimann ◽  
Christian Paulsen ◽  
Jutta Kösters ◽  
...  

Abstract The scandium-rich intermetallic compounds Sc50T13In3 (T = Ni, Ru, Pd) were synthesized from the elements in sealed tantalum crucibles in an induction furnace. The samples were studied through Guinier powder patterns and their structures were refined from single-crystal X-ray diffractometer data. The Sc50T13In3 phases are site occupancy (coloring) variants of the aristotype Ag7+xMg26–x (Fm$$ \bar{3} $$ 3 ¯ , cF264). Refinements of the occupancy parameters indicated one mixed occupied site for each crystal, leading to the refined compositions Sc50Ni13.16(1)In2.84(1), Sc49.59(1)Ru13In3.41(1), and Sc50Pd13.65(2)In2.35(2). The complex structures can be explained by a condensation of cubes (CN 8), sphenocorona (CN 10), and icosahedra (CN 12). The samples with nickel and palladium are Pauli paramagnets. Graphic abstract


2019 ◽  
Vol 86 (5) ◽  
Author(s):  
Jie Liu ◽  
Lorenz Adrian ◽  
Max M. Häggblom

ABSTRACT Organohalide respiration is an important process in the global halogen cycle and for bioremediation. In this study, we compared the global transcriptomic and proteomic analyses of Desulfoluna spongiiphila strain AA1, an organohalide-respiring member of the Desulfobacterota isolated from a marine sponge, with 2,6-dibromophenol or with sulfate as an electron acceptor. The most significant difference of the transcriptomic analysis was the expression of one reductive dehalogenase gene cluster (rdh16), which was significantly upregulated with the addition of 2,6-dibromophenol. The corresponding protein, reductive dehalogenase RdhA16032, was detected in the proteome under treatment with 2,6-dibromophenol but not with sulfate only. There was no significant difference in corrinoid biosynthesis gene expression levels between the two treatments, indicating that the production of corrinoid in D. spongiiphila is constitutive or not specific for organohalide versus sulfate respiration. Electron-transporting proteins or mediators unique for reductive dehalogenation were not revealed in our analysis, and we hypothesize that reductive dehalogenation may share an electron-transporting system with sulfate reduction. The metabolism of D. spongiiphila, predicted from transcriptomic and proteomic results, demonstrates high metabolic versatility and provides insights into the survival strategies of a marine sponge symbiont in an environment rich in organohalide compounds and other secondary metabolites. IMPORTANCE Respiratory reductive dehalogenation is an important process in the overall cycling of both anthropogenic and natural organohalide compounds. Marine sponges produce a vast array of bioactive compounds as secondary metabolites, including diverse halogenated compounds that may enrich for dehalogenating bacteria. Desulfoluna spongiiphila strain AA1 was originally enriched and isolated from the marine sponge Aplysina aerophoba and can grow with both brominated compounds and sulfate as electron acceptors for respiration. An understanding of the overall gene expression and the protein production profile in response to organohalides is needed to identify the full complement of genes or enzymes involved in organohalide respiration. Elucidating the metabolic capacity of this sponge-associated bacterium lays the foundation for understanding how dehalogenating bacteria may control the fate of organohalide compounds in sponges and their role in a symbiotic organobromine cycle.


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