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Human cytochrome P450 (P450) 27A1 catalyzes the hydroxylation of cholesterol and vitamin D derivatives. P450 27A1 is localized in the mitochondria and is reduced by its redox partner protein adrenodoxin twice for each catalytic cycle. The reliance on adrenodoxin is conserved across all human mitochondrial P450 enzymes. This study examines the adrenodoxin interaction with P450 27A1 and draws comparisons with studies of other P450 enzymes to determine if differences exist. The P450-adrenodoxin complex structure was examined by chemical crosslinking and analyzed by mass spectrometry. The effect of adrenodoxin concentration on P450 27A1 function was assessed by studying effects on steady state enzyme kinetics parameters and equilibrium substrate binding. The results suggest that adrenodoxin binds to P450 27A1 at a proximal site like other P450 enzymes but differs in the specific residues involved. Furthermore, the presence of adrenodoxin and/or substrate decreases the number of interprotein and intraprotein crosslinks observed, indicating that these components change the conformation of the P450 enzyme. Increased adrenodoxin concentration causes the P450 and vitamin D3 kcat value to increase, the kcat/Km value to decrease, and the substrate Kd to remain constant. These results suggest adrenodoxin alters enzyme efficiency beyond electron transfer without affecting substrate loading. The adrenodoxin effects on P450 27A1 kinetics and equilibrium constants differ from those of other human mitochondrial P450 enzymes. In total, these structural and functional studies suggest that while the general adrenodoxin binding site and function is conserved across P450 enzymes, the details and additional effects of this interaction vary. 

Abstract To understand how behaviors arise in animals, it is necessary to investigate both the neural circuits and the biomechanics of the periphery. A tractable model system for studying multifunctional control is the feeding apparatus of the marine molluskAplysia californica. Previousin silicoandin robotomodels have investigated how the nervous and muscular systems interact in this system. However, these models are still limited in their ability to matchin vivodata both qualitatively and quantitatively. We introduce a new neuromechanical model ofAplysiafeeding that combines a modified version of a previously developed neural model with a novel biomechanical model that better reflects the anatomy and kinematics ofAplysiafeeding. The model was calibrated using a combination of previously measured biomechanical parameters and hand-tuning to behavioral data. Using this model, simulated feeding experiments were conducted, and the resulting behavioral metrics were compared to animal data. The model successfully produces three key behaviors seen inAplysiaand demonstrates a good quantitative agreement with biting and swallowing behaviors. Additional work is needed to match rejection behavior quantitatively and to reflect qualitative observations related to the relative contributions of two key muscles, the hinge and I3. Future improvements will focus on incorporating the effects of deformable 3D structures in the simulated buccal mass. 

Ten new ruthenium compounds based on the N,N,N,N-chelate Me2bpbMe2 (bpb = 1,2-bis(pyridine-2-carboximido)benzene) have prepared and characterized by 1H NMR and IR spectroscopy. The monocarbonyl compound (Me2bpbMe2)Ru(CO)(H2O) compound was generated from the reaction of the free base Me2bpbMe2H2 with Ru3(CO)12 in refluxing DMF. Isoamyl nitrite reacts with this compound to yield the trans-addition nitrosyl alkoxide (Me2bpbMe2)Ru(NO)(O-i-C5H11). Nitrosothiols similarly add in a formal trans-addition manner to yield (Me2bpbMe2)Ru(NO)(SR/Ar) (SR/Ar = S-i-C5H11, SPh, SC6F4H, SC(Me)2CHNHC(O)Me) derivatives. The (Me2bpbMe2)Ru(NO)(O-i-C5H11) compound undergoes alkoxide exchange reactions with PhOH and HOC6F4H to generate (Me2bpbMe2)Ru(NO)(OPh) and (Me2bpbMe2)Ru(NO)(OC6F4H), respectively. The neutral alkoxide/aryloxide nitrosyl compounds exhibit higher NO bands (1809–1842 cm-1) relative to their thiolate analogues (1755–1823 cm-1). The X-ray crystal structures of (Me2bpbMe2)Ru(NO)(OPh), (Me2bpbMe2)Ru(NO)(OC6F4H), and (Me2bpbMe2)Ru(NO)(SPh), have been determined, and reveal linear axial (O)N–Ru–O/S linkages consistent with trans positioning of the NO and aryloxide and -thiolate groups, and near-linear Ru–N–O moieties (164–174°) consistent with these complexes being formulated as {RuNO}6 species. The electrooxidation behavior of (Me2bpbMe2)Ru(NO)(OC6F4H), (Me2bpbMe2)Ru(NO)(SC6F4H), and (Me2bpbMe2)Ru(NO)(SPh) were examined by cyclic voltammetry and IR spectroelectrochemistry in CH2Cl2. (Me2bpbMe2)Ru(NO)(OC6F4H) and (Me2bpbMe2)Ru(NO)(SC6F4H) display reversible first oxidations, whereas (Me2bpbMe2)Ru(NO)(SPh) displays an irreversible first oxidation with likely loss of the thiolate ligand. Chemical reactivity of (Me2bpbMe2)Ru(NO)(SPh) with H+ and Me+ results in the generation of the free thiol PhSH and thioether PhSMe, respectively. 

MicroRNAs (miRNAs) are small non-protein-coding RNAs that regulate gene expression in many eukaryotes. Next-generation sequencing of small RNAs (small RNA-seq) is central to the discovery and annotation of miRNAs. Newly annotated miRNAs and their longer precursors encoded byMIRNAloci are typically submitted to databases such as the miRBase microRNA registry following the publication of a peer-reviewed study. However, genome-wide scans using small RNA-seq data often yield high rates of false-positiveMIRNAannotations, highlighting the need for more robust validation methods. miRScore was developed as an independent and efficient tool for evaluating newMIRNAannotations using sRNA-seq data. miRScore combines structural and expression-based analyses to provide rapid and reliable validation of newMIRNAannotations. By providing users with detailed metrics and visualization, miRScore enhances the ability to assess confidence inMIRNAannotations. miRScore has the potential to advance the overall quality ofMIRNAannotations by improving accuracy of new submissions to miRNA databases and serving as a resource for re-evaluating existing annotations.