A5010882529- Inflammatory mediators and NSAID effects
- Estrogen and related hormone effects
- Eicosanoids and Hypertension Pharmacology
- Steroid Chemistry and Biochemistry
- Asthma and respiratory diseases
- Pharmacogenetics and Drug Metabolism
- Mass Spectrometry Techniques and Applications
- Synthesis of β-Lactam Compounds
- bioluminescence and chemiluminescence research
- Peptidase Inhibition and Analysis
- Receptor Mechanisms and Signaling
- Neuropeptides and Animal Physiology
- Nitric Oxide and Endothelin Effects
- Synthesis and Characterization of Heterocyclic Compounds
- Chemical Synthesis and Analysis
- Electron Spin Resonance Studies
- Computational Drug Discovery Methods
- Viral Infectious Diseases and Gene Expression in Insects
- Crystal structures of chemical compounds
- Protein Structure and Dynamics
- Pediatric health and respiratory diseases
- Vitamin K Research Studies
- Chemical Reactions and Isotopes
- Various Chemistry Research Topics
- Ion channel regulation and function
Donald Danforth Plant Science Center
2014-2016
Agricultural Research Service
2014-2016
United States Department of Agriculture
2016
Pfizer (United States)
2005-2010
Monsanto (United States)
1987-2002
Vanderbilt University
1997
University of Chicago
1996
University of Colorado Anschutz Medical Campus
1994
Hafslund (Norway)
1992
University of Colorado Health
1991-1992
The enzyme cyclo-oxygenase catalyses the oxygenation of arachidonic acid, leading to formation prostaglandins. Recently two forms have been described: a constitutive (COX-1) present in most cells and tissues, an inducible (COX-2) isoenzyme observed many response pro-inflammatory cytokines. Constitutive human (hCOX-1 hCOX-2) were cloned expressed insect cells, utilizing baculovirus expression system. hCOX-1 had specific activity 18.8 mumol O2/mg with Km 13.8 microM for arachidonate Vmax. 1500...
Nonsteroidal anti-inflammatory drugs (NSAIDs) currently available for clinical use inhibit both COX-1 and COX-2. This suggests that clinically useful NSAIDs pro-inflammatory prostaglandins (PGs) derived from the activity of COX-2, as well PGs in tissues like stomach kidney (via COX-1). A new class compounds has recently been developed (SC-58125) have a high degree selectivity inducible form cyxlooxygenase (COX-2) over constitutive (COX-1). unique exhibit time-dependent irreversible...
In Escherichia coli high-level production of some heterologous proteins (specifically, human prorenin, renin, and bovine insulin-like growth factor 2) resulted in the induction two new E. heat shock proteins, both which have molecular masses 16 kDa are tightly associated with inclusion bodies formed during protein production. We named these body-associated IbpA IbpB. The coding sequences for IbpB were identified isolated from Kohara gene bank. genes (ibpA ibpB) located at 82.5 min on...
Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit the formation of prostaglandins by cyclo-oxygenases (COX). The discovery a second COX isoform (COX-2) associated with inflammation led to agents that selectively COX-2, e.g. celecoxib. We evaluated kinetics inhibition celecoxib and several NSAIDs. Celecoxib displays classic competitive on COX-1 (Ki = 10-16 μM). An initial interaction COX-2 can also be discerned 11-15 μM), followed time-dependent leading potent inhibition, characterized...
Autotaxin is the enzyme responsible for production of lysophosphatidic acid (LPA) from lysophosphatidyl choline (LPC), and it up-regulated in many inflammatory conditions, including but not limited to cancer, arthritis, multiple sclerosis. LPA signaling causes angiogenesis, mitosis, cell proliferation, cytokine secretion. Inhibition autotaxin may have anti-inflammatory properties a variety diseases; however, this hypothesis has been tested pharmacologically because lack potent inhibitors....
The determination of protein nitrotyrosine content has become a frequently used technique for the detection oxidative tissue damage. Protein nitration been suggested to be final product production highly reactive nitrogen oxide intermediates (<i>e.g.</i> peroxynitrite) formed in reactions between nitric (NO<sup>⋅</sup>) and oxygen-derived species such as superoxide. enzyme prostaglandin H synthase-2 (PHS-2) forms one or more tyrosyl radicals during its enzymatic catalysis formation. In...
Abstract Leukotriene-A4 hydrolase (EC 3.3.2.6) cleaved the NH2-terminal amino acid from several tripeptides, typified by arginyl-glycyl-aspartic acid, arginyl-glycyl-glycine, and arginyl-histidyl-phenylalanine, with catalytic efficiencies (kcat/Km) > or = 1 x 10(6) M-1 s-1. This exceeds 10-fold kcat/Km for its lipid substrate leukotriene A4. Catalytic efficiency declined dipeptides which had ratios 10-100-fold lower than tripeptides. Tetrapeptides pentapeptides were even poorer substrates...
Captopril((2S)-l-(3-mercapto-2-methyl-propionyl)-L-proline) inhibited the bifunctional, Zn2+-containing
The two isoforms of cyclooxygenase, COX-1 and COX-2, are acetylated by aspirin at Ser-530 Ser-516, respectively, in the cyclooxygenase active site. Acetylated COX-2 is essentially a lipoxygenase, making 15-(<i>R</i>)-hydroxyeicosatetraenoic acid (15-HETE) 11-(<i>R</i>)-hydroxyeicosatetraenoic (11-HETE), whereas unable to oxidize arachidonic any products. Because COX structurally similar share approximately 60% amino identity, we postulated that differences within sites must account for...
The discovery of a second isoform cyclooxygenase (COX) led to the search for compounds that could selectively inhibit COX-2 in humans while sparing prostaglandin formation from COX-1. Celecoxib and rofecoxib were among molecules developed these efforts. We report here pharmacological properties third selective inhibitor, valdecoxib, which is most potent vitro marketed inhibitors we have studied. Recombinant human COX-1 used screen new highly compare kinetic mechanisms binding enzyme...
The metabolism of atriopeptin prohormone ANF1-126 was examined with the aid two separate radioimmunoassays, one detecting C-terminal atriopeptins and other a fragment N-terminus. Intact standards are recognized in both assays, whereas only detected by assay. Both N-terminal immunoreactivities were rat plasma simultaneously elevated following intravenous administration desamino-arginine-vasopressin. Atriopeptin immunoreactivity returned to basal levels within 60 min after desamino-arginine...
C4 photosynthesis in grasses requires the coordinated movement of metabolites through two specialized leaf cell types, mesophyll (M) and bundle sheath (BS), to concentrate CO2 around Rubisco. Despite importance transporters this process, few have been identified or rigorously characterized. In maize (Zea mays), DCT2 has proposed function as a plastid-localized malate transporter is preferentially expressed BS cells. Here, we characterized role leaves using Activator-tagged mutant alleles....
Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit the formation of prostaglandins by cyclo-oxygenases (COX). The discovery a second COX isoform (COX-2) associated with inflammation led to agents that selectively COX-2, e.g. celecoxib. We evaluated kinetics inhibition celecoxib and several NSAIDs. Celecoxib displays classic competitive on COX-1 (Ki = 10-16 μM). An initial interaction COX-2 can also be discerned 11-15 μM), followed time-dependent leading potent inhibition, characterized...
Cyclo-oxygenase (COX) enzymes are the targets for non-steroidal anti-inflammatory drugs (NSAIDs). These demonstrate a variety of inhibitory mechanisms, which include simple competitive, as well slow binding and irreversible inhibition. In general, most NSAIDs inhibit COX-1 -2 by similar mechanisms. A unique class diarylheterocyclic inhibitors has been developed that is highly selective COX-2 virtue distinct mechanisms each isoenzyme. Several these inhibitors, with varying selectivity, have...
Site-directed mutants of prostaglandin-endoperoxide synthase-2 (PGHS-2) with changes in the peroxidase active site were prepared by mutagenesis, expressed Sf-9 cells, and purified to homogeneity. The distal histidine, His193, was mutated alanine glutamine, Gln189, changed asparagine, valine, arginine. guaiacol activities H193A, Q189V, Q189R drastically reduced levels observed absence protein; only Q189N retained wild-type PGHS-2 (wtPGHS-2) activity. mechanism hydroperoxide reduction...
Abstract Suicide inactivation of leukotriene (LT) A4 hydrolase/aminopeptidase occurs via an irreversible mechanism-based process which is saturable, pseudo firstorder, and dependent upon catalysis. Data obtained with either recombinant enzyme or purified from human leukocytes were similar. Apparent binding constants rate are equivalent, compatible a single type substrate-enzyme complex partitions between two fates, turnover inactivation. Both catalytic functions inactivated, consistent...
Leukotriene B(4) (LTB(4)) is a pro-inflammatory mediator that has been implicated in the pathogenesis of number diseases including inflammatory bowel disease (IBD) and psoriasis. Since action LTA(4) hydrolase rate-limiting step for LTB(4) production, this enzyme represents an attractive pharmacological target suppression production. From in-house screening program, SC-22716 (1, 1-[2-(4-phenylphenoxy)ethyl]pyrrolidine) was identified as potent inhibitor hydrolase. Structure-activity...
Two compounds (celecoxib and valdecoxib) from the diarylheterocycle class of cyclooxygenase inhibitors were radiolabeled used to characterize their binding cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), several single-point variants COX-2 (Val523Ile, Tyr355Ala, Arg120Ala, Arg120Gln, Arg120Asn) one triple-point variant [Val523Ile, Arg513His, Val434Ile (IHI)]. We demonstrate highly specific saturable these COX-2. Under same assay conditions, little or no COX-1 could be detected. The...
Cyclo-oxygenase (COX) enzymes are the targets for non-steroidal anti-inflammatory drugs (NSAIDs). These demonstrate a variety of inhibitory mechanisms, which include simple competitive, as well slow binding and irreversible inhibition. In general, most NSAIDs inhibit COX-1 −2 by similar mechanisms. A unique class diarylheterocyclic inhibitors has been developed that is highly selective COX-2 virtue distinct mechanisms each isoenzyme. Several these inhibitors, with varying selectivity, have...