Poly(ADP-ribose) Catabolism Triggers AMP-dependent Mitochondrial Energy Failure

Upon massive DNA damage, hyperactivation of the nuclear enzyme poly(ADP-ribose) polymerase (PARP)-1 causes severe depletion of intracellular NAD and ATP pools as well as mitochondrial dysfunction. Thus far, the molecular mechanisms contributing to PARP-1-dependent impairment of mitochondrial functioning have not been identified. We found that degradation of the PARP-1 product poly(ADP-ribose) through the concerted actions … Continued

In Vitro Characterization of a Recombinant Blh Protein from an Uncultured Marine Bacterium as a β-carotene 15,15'-dioxygenase

Codon optimization was used to synthesize the blh gene from the uncultured marine bacterium 66A03 for expression in Escherichia coli. The expressed enzyme cleaved β-carotene at its central double bond (15,15′) to yield two molecules of all-trans-retinal. The molecular mass of the native purified enzyme was approximately 64 kDa as a dimer of 32-kDa subunits. … Continued

LysM Domains from Pteris Ryukyuensis Chitinase-A: A Stability Study and Characterization of the Chitin-binding Site

The LysM domain probably binds peptidoglycans, but how it does so has yet to be described. For this report, we measured the thermal stabilities of recombinant LysM domains derived from Pteris ryukyuensis chitinase-A (PrChi-A) and monitored their binding to N-acetylglucosamine oligomers ((GlcNAc)n) using differential scanning calorimetry, isothermal titration calorimetry, and NMR spectroscopy. We thereby characterized … Continued

CYP2E1 Substrate Inhibition: Mechanistic Interpretation Through an Effector Site for Monocyclic Compounds

In this study we offer a mechanistic interpretation of the previously known but unexplained substrate inhibition observed for CYP2E1. At low substrate concentrations, p-nitrophenol (pNP) was rapidly turned over (47 min-1) with relatively low Km (24 µM); nevertheless, at concentrations of >100 µM, the rate of pNP oxidation gradually decreased as a second molecule bound … Continued

The Structure of the R184A Mutant of the Inositol Monophosphatase Encoded by SuhB and Implications for Its Functional Interactions in Escherichia Coli

The Escherichia coli product of the suhB gene, SuhB, is an inositol monophosphatase (IMPase) that is best known as a suppressor of temperature-sensitive growth phenotypes in E. coli. To gain insights into these biological diverse effects, we determined the structure of the SuhB R184A mutant protein. The structure showed a dimer organization similar to other … Continued

Critical Role for Polar Residues in Coupling Leukotriene B4 Binding to Signal Transduction in BLT1

Leukotriene B(4) (LTB(4)) mediates a variety of inflammatory diseases such as asthma, arthritis, atherosclerosis, and cancer through activation of the G-protein-coupled receptor, BLT1. Using in silico molecular dynamics simulations combined with site-directed mutagenesis we characterized the ligand binding site and activation mechanism for BLT1. Mutation of residues predicted as potential ligand contact points in transmembrane … Continued

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