40 research outputs found
Yeast squalene synthase. A mechanism for addition of substrates and activation by NADPH
Journal ArticleSqualene synthase catalyzes the condensation of two molecules of farnesyl diphosphate (FPP) to give presqualene diphosphate (PSPP) and the subsequent reductive rearrangement of PSPP to squalene. Previous studies of the mechanism of addition of FPP to the enzyme have led to conflicting interpretations of initial velocity measurements (Beytia, E., Qureshi, A. A., and Porter, J.W. (1973) J. Biol. Chem. 248, 1856-1867; Agnew, W.S., and Popjak, G. (1978) J. Biol. Chem. 253, 4566-4573). Initial velocities for synthesis of PSPP and squalene were measured over a wider range of FPP and NADPH concentrations than previously reported, using a soluble form of recombinant enzyme. In the absence of NADPH, PSPP formation was activated by FPP at concentrations above approximately 0.5 microM. At fixed levels of NADPH, the dependence of initial rates of PSPP and squalene synthesis on FPP concentrations indicated that the C15 substrate added by a sequential mechanism. In addition, NADPH stimulated synthesis of PSPP by 40-fold at saturating levels of the cofactor. This stimulation is, at least in part, by reduction of PSPP to squalene
Utilization of marine industry waste derived collagen hydrolysate as peroxide inhibition agents in lipid‐based food
BRAF and AXL oncogenes drive RIPK3 expression loss in cancer.
Necroptosis is a lytic programmed cell death mediated by the RIPK1-RIPK3-MLKL pathway. The loss of Receptor-interacting serine/threonine-protein kinase 3 (RIPK3) expression and necroptotic potential have been previously reported in several cancer cell lines; however, the extent of this loss across cancer types, as well as its mutational drivers, were unknown. Here, we show that RIPK3 expression loss occurs progressively during tumor growth both in patient tumor biopsies and tumor xenograft models. Using a cell-based necroptosis sensitivity screen of 941 cancer cell lines, we find that escape from necroptosis is prevalent across cancer types, with an incidence rate of 83%. Genome-wide bioinformatics analysis of this differential necroptosis sensitivity data in the context of differential gene expression and mutation data across the cell lines identified various factors that correlate with resistance to necroptosis and loss of RIPK3 expression, including oncogenes BRAF and AXL. Inhibition of these oncogenes can rescue the RIPK3 expression loss and regain of necroptosis sensitivity. This genome-wide analysis also identifies that the loss of RIPK3 expression is the primary factor correlating with escape from necroptosis. Thus, we conclude that necroptosis resistance of cancer cells is common and is oncogene driven, suggesting that escape from necroptosis could be a potential hallmark of cancer, similar to escape from apoptosis
Preparation and reconstitution with divalent metal ions of class I and class II Clostridium histolyticum apocollagenases
Discovery of a Potent Inhibitor Class with High Selectivity toward Clostridial Collagenases
Characterization of 58-kilodalton human neutrophil collagenase: comparison with human fibroblast collagenase
Orally Active Squalene Synthase Inhibitors: Bis((acyloxy)alkyl) Prodrugs of the α-Phosphonosulfonic Acid Moiety
Activation of human neutrophil procollagenase by stromelysin 2
Knauper V, Murphy G, Tschesche H. Activation of human neutrophil procollagenase by stromelysin 2. EUROPEAN JOURNAL OF BIOCHEMISTRY. 1996;235(1-2):187-191.Neutrophil procollagenase (MMP-8) was efficiently activated by incubation with active stromelysin 2 (MMP-10). A single-step activation mechanism involving the cleavage of the Gly78-Phe79 peptide bond at the end of the propeptide domain was observed. Determination of the collagenolytic activity revealed the generation of active neutrophil collagenase displaying high specific activity. When compared with the specific activity following mercurial activation, which generates active collagenase by autoproteolytic cleavage of either Phe79-Met80 or Met80-Leu81 peptide bonds [Blaser, J., Knauper, V., Osthues, A., Reinke, H. & Tschesche, H. (1991) fur: J. Biochem. 202, 1223-1230], the specific activity of the stromelysin-2-activated enzyme was considerably higher. Thus, human neutrophil procollagenase was 'superactivated' by stromelysin 2, as was recently shown for the stromelysin-1-activated enzyme [Knauper, V., Wilhelm, S. M., Seperack, P. K., De Clerck,Y. A., Langley, K. E., Osthues, A. & Tschesche, H. (1993 a) Biochem. J. 295, 581-586]