Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P00790 (PGA)
 2,475 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Ribulose-diphosphate carboxylase from Thiobacillus novellus has been purified to hemogeneity as observed by polyacrylamide gel electrophoresis and U.V. light observation during sedimentation velocity analysis. The optimum pH for the enzyme with Tris-HCl buffers was about 8.2. Concentrations of this buffer in excess of 80 mM were inhibitory. The apparent Km for RuDP was about 14.8 muM with a Hill value of 1.5, for HCO3- the apparent Km was about 11.7 mM with an n value of 1.18 and for Mg2+ about 0.61 mM. The enzyme was specific for this cation. Relatively high concentrations of either Hg2+ or pCMB were required before significant inhibition was observed. Activity declined slowly during a 4-hr incubation period in either 3.0 M or 8.0 M urea. Incubation for 12 hrs resulted in complete loss of activity which was not prevented by 10 mM Mg2+ and was not reversed by dialysis and subsequent addition of 10 mM cysteine. Polyacrylamide gel electrophoresis revealed a loss of the major band and the appearance of 2 new bands. SDS polyacrylamide gel electrophoresis gave an average M.W. of 73500 +/- 2500 for the slower moving band and 12250 +/- 2500 for the faster moving. However, incubation in urea for up to 40 hrs revealed a decrease in the M.W. of the slower moving band to about 60000. The Ea for the enzyme was calculated to be about 18.85 kcal mole-1, with the possibility of a "break" between 40 and 50 degrees C. The Q10 was 3.07 between 20 and 30 degrees C whereas between 30 to 40 degrees C it was 3.31. Only phosphorylated compounds caused significant inhibition of enzyme activity. They included ADP, FDP, F6P, G6P, PEP, 6PG, 2-PGA, R1P, R5P, and Ru5p.
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PMID:Properties and regulation of ribulose diphosphate carboxylase from Thiobacillus novellus. 24 94

Human nucleus pulposus and annulus fibrosus, obtained at autopsy from patients 7-30 years of age, were extracted with 2 M guanidine-HCl (pH 5.82) to remove proteoglycans, then stirred with pepsin in 0.5 M acetic acid, followed by three 24-h extractions with 1 M NaCl (pH 7.5) and one 24-h extraction with 2 M KSCN (potassium thiocyanate) (pH 7.2). Pepsin and NaCl solubilized an average of about 30% of nucleus pulposus collagen and 18% of annulus fibrosus collagen. KSCN extracted a further 34% of nucleus pulposus collagen and only 4% of annulus fibrosus collagen. CM-cellulose chromatography of nucleus and annulus collagen purified from the pepsin, NaCl and KSCN supernatants consistently revealed only one peak, always appearing slightly ahead of the alpha1 position for rat tail tendon type I collagen. Polyacrylamide and SDS-gel electrophoresis consistently revealed only one band with the mobility of alpha1 chains. Amino acid composition of collagen from nucleus and annulus is comparable to those of mammalian and avian cartilage type II collagen, and distinctly different from those of rat tail tendonand guinea pig skin type I collagens. Periodate oxidation of nucleus and annulus collagens showed that 81% and 67%, respectively, of the hydroxylysine residues survive treatment, compared to 71% for bovine articular cartilage collagen and 17% for guinea pig skin collagen. Total hexose analysis revealed 1.8 muM and 2.0 muM hexose per muM periodate-stable hydroxylysine in nucleus and annulus collagens, respectively. Ion exchange chromatography showed the presence of glucose and galactose in a ratio of 0.92:1 in nucleas collagen and 1.07:1 in annulus collagen. Pepsin-solubilized, NaCl-extracted collagen from nucleus and annulus formed native-type fibrils in vitro. The banding patterns of ATP-induced segment-long-spacing precipitates of nucleus and annulus collagens were identical to each other and indistinguishable from those of cartilage (type II) collagen, but distinctly different from those of rat tail tendon (type I) collagen. These data suggest that the collagen which can be extracted after limited pepsin attack of human nucleus and annulus is of the form [alpha1 (II)]3.
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PMID:Pepsin-solubilized collagen of human nucleus pulposus and annulus fibrosus. 78 25

Cyclopentenone prostaglandins (PGs) such as delta 12-PGJ2 and PGA are potent inducers of growth inhibition in a variety of cultured cells, including epidermal cells. These PGs are actively transported into cells by a specific carrier on cell membrane and accumulate in cell nuclei with binding to nuclear protein. To clarify the mechanism of cytotoxicity of these PGs in epidermal cells, we examined the effects of delta 12-PGJ2 on protein synthesis and cytoskeleton in the PAM 212 transformed mouse epidermal cell line. Cycloheximide at 1 microgram/ml culture medium exhibited a protective effect on cell growth inhibition of PAM 212 cells by delta 12-PGJ2. The analysis of cell lysate protein patterns by SDS-polyacrylamide gel electrophoresis revealed that 12-h incubation with delta 12-PGJ2 increased the amount of 70 kD protein in PAM 212 cells. The amount of 70 kD protein in delta 12-PGJ2-treated cells was markedly decreased by cotreatment with cycloheximide. This 70 kD protein was also induced in PAM 212 cells with treatment at 43 degrees C for 90 min, indicating that this synthesized protein belongs to the heat shock protein. The addition of delta 12-PGJ2 to confluent PAM 212 cells resulted in the disappearance of action filament, as visualized by fluorescent labeled phallacidine, but in contrast, keratin filament appeared to be intact during 12-h incubation with delta 12-PGJ2 at a concentration of 5 micrograms/ml culture medium. These results suggest that the cytotoxicity of cyclopentenone PGs is at least in part due to induction of the synthesis of some protein(s), probably one of the heat shock proteins, and the damage to the actin filament in transformed cultured epidermal cells.
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PMID:Effects of cytotoxic prostaglandin, delta 12-PGJ2 on protein synthesis and cytoskeleton in transformed epidermal cells in culture. 169 39

An acid proteinase of Dirofilaria immitis worms was purified 437-fold by gel filtration on Sephadex G-75 followed by pepstatin-Agarose gel affinity chromatography. The enzyme with a molecular weight of 42 kDa was homogeneous as judged by both affinity chromatography and SDS-polyacrylamide gel electrophoresis. Polyacrylamide disc electrophoresis at pH 8.9, however, revealed that the enzyme was composed of five multi-forms, all carrying proteinase activity. Optimum pH of the enzyme was in the range of pH 2.8 to 3.4, and its isoelectric point ranged between 5.8 and 6.4. The purified proteinase showed a potent activity against hemoglobin and myoglobin releasing acid soluble peptides, but not free amino acids. When enzymatic properties of the proteinase was compared with mammalian cathepsin D and pepsin, D. immitis proteinase activity was reduced to about 80% of the initial activity by incubating at neutral pH and 50 degrees C for 5 min, just like cathepsin D, which remained intact. Pepsin activity was completely destroyed under the same condition. An aspartic proteinase inhibitor, 1,2-epoxy-3-(p-nitrophenoxy)propane, which inhibited pepsin by 30% at 37 degrees C for 10 min, did show little effects on D. immitis proteinase and cathepsin D. Inhibitory effect of diazoacetyl-DL-norleucine methyl ester (DAN) on D. immitis proteinase was intermediate (50% after 60 min). Immunolocalization of the proteinase in the worm tissue using its monoclonal antibodies revealed that the enzyme was localized in the intestine as well as uterine wall and some small granules of microfilariae in the uterus.
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PMID:Purification and characterization of an acid proteinase from Dirofilaria immitis worms. 854 Mar 32