Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0019204 (hepatocellular carcinoma)
 71,386 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Glutamine-dependent carbamyl phosphate synthetase [EC 2.7.2.9] was purified 1,300-fold from rat ascites hepatoma cells (AH 13) as a multienzyme complex with aspartate transcarbamylase[EC 2.1.3.2] and dihydroorotase[EC 3.5.2.3], using dimethyl sulfoxide, glycerol, and dithiothreitol as stabilizers. The purified complex was essentially homogeneous on agarose-acrylamide composite gel electrophoresis and analytical ultracentrifugation. Its molecular weight was estimated to be about 870,000 by sedimentation equilibrium studies. After alkylation with iodoacetamide or reduction with 0.6% dithiothreitol at 100 degrees, the complex gave a single band on polyacrylamide gel electrophoresis in sodium dodecyl sulfate in a position corresponding to a molecular weight of 210,000. These results indicate that the complex consists of four subunits of similar size.
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PMID:Purification of homogeneous glutamine-dependent carbamyl phosphate synthetase from ascites hepatoma cells as a complex with aspartate transcarbamylase and dihydroorotase. 17 92

Carbamoyl-phosphate synthetase II [EC 6.3.5.5] of rat ascites hepatoma cells (AH 13), the first and regulatory enzyme of de novo pyrimidine nucleotide biosynthesis, exists as a multienzyme complex (molecular weight, 870,000) with aspartate carbamoyltransferase [EC 2.1.3.2] and dihydroorotase [EC 3.5.2.3] (Mori, M. & Tatibana, M. (1975) J. Biochem. 78, 239-242). The purified complex was phosphorylated by the catalytic subunit of cAMP-dependent protein kinase [EC 2.7.1.37] of rabbit skeletal muscle. The incorporation of 32Pi was 2.2 mol/mol of the complex. The phosphorylation was completely inhibited by the inhibitor protein of the cAMP-dependent protein kinase. Among the substrates and effectors of the enzyme complex tested, only MgUTP, an allosteric inhibitor of carbamoyl-phosphate synthetase II, strongly inhibited the phosphorylation; this inhibition was due probably to the competition of MgUTP with y inhibited by the inhibitor protein of the cAMP-dependent protein kinase. Among the substrates and effectors of the enzyme complex tested, only MgUTP, an allosteric inhibitor of carbamoyl-phosphate synthetase II, strongly inhibited the phosphorylation; this inhibition was due probably to the competition of MgUTP with y inhibited by the inhibitor protein of the cAMP-dependent protein kinase. Among the substrates and effectors of the enzyme complex tested, only MgUTP, an allosteric inhibitor of carbamoyl-phosphate synthetase II, strongly inhibited the phosphorylation; this inhibition was due probably to the competition of MgUTP with the substrate MgATP for the protein kinase. The complex that was phosphorylated by cAMP-dependent protein kinase was dephosphorylated by phosphoprotein phosphatase [EC 3.1.3.16] of rat skeletal muscle. The complex was also phosphorylated by cAMP-independent protein kinase activity present in the extract of AH 13 cells and dephosphorylated by phosphoprotein phosphatase activity of the same origin. These results suggest that the complex is subject to phosphorylation and dephosphorylation in the living cells. Phosphorylation of the complex by cAMP-dependent protein kinase was associated only with a slight change, albeit definite, in the activity of carbamoyl-phosphate synthetase II under the assay conditions. Thus, the physiological significance of phosphorylation-dephosphorylation remains to be further studied.
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PMID:Phosphorylation and dephosphorylation of carbamoyl-phosphate synthetase II complex of rat ascites hepatoma cells. 611 55

Carbamoyl-phosphate synthetase II of higher animals, the first enzyme of de novo pyrimidine biosynthesis, forms a multienzyme complex with aspartate carbamoyltransferase and dihydroorotase, the second and third enzymes of the pathway. The hypothesis that the complex serves to channel carbamoyl-phosphate, synthesized by the first enzyme of the complex, to the second enzyme was tested using a highly purified complex preparation from Yoshida ascites hepatoma cells (AH 13). Experimentally, aspartate carbamoyltransferase in the complex was allowed to compete with exogenously added ornithine carbamoyltransferase, another carbamoyl-phosphate-utilizing enzyme, for carbamoyl-phosphate which was either synthesized endogenously or added exogenously. The ratios of amounts of the two enzymic products, carbamoyl-aspartate and citrulline, were compared. In the absence of enzyme stabilizers dimethyl sulfoxide or glycerol, a slight channeling of the intermediate in the complex was observed. The further addition of 5-phosphoribosyl 1-pyrophosphate, MgUTP (positive and negative allosteric effectors of carbamoyl-phosphate synthetase II), 30% (v/v) dimethyl sulfoxide or 30% (w/v) glycerol did not affect the extent of channeling. It was slightly increased in the presence of 7.5% (v/v) dimethyl sulfoxide plus 2.5% (w/v) glycerol. Any shift of the assay temperature, pH or concentration of MgATP or of the enzyme complex resulted in little further increase in the extent of channeling. Even when a larger amount of the enzyme complex was used to approximate physiological conditions, there was no increase in the extent of channeling either without or with allosteric effectors. MgUTP even abolished channeling under these conditions. These results indicate that carbamoyl-phosphate can be channeled in the multienzyme complex of AH 13 cells, but the extent of channeling is very small, contrary to expectation.
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PMID:Studies on channeling of carbamoyl-phosphate in the multienzyme complex that initiates pyrimidine biosynthesis in rat ascites hepatoma cells. 613 83

Genomic stability was investigated in Chinese hamster ovary (CHO) and human hepatocellular carcinoma HepG2 cells selected for growth in the presence of cytotoxic concentrations of N-(phosphonacetyl-L-asparate) (PALA). In CHO cells selected with 9 x LD50 PALA the carbamyl p-synthetase, aspartate transcarbamylase and dihydroorotase (CAD) gene complex was amplified two-fold while in HepG2 cells selected at comparable PALA concentrations a 7- to 10-fold increase in the CAD gene was observed. Concomitant with amplification of the CAD gene were increases in CAD mRNA and protein expression in both CHO and HepG2 cells. In long-term cultures of HepG2 cells the CAD gene underwent spontaneous amplification (5-fold) in the absence of PALA treatment with increasing passage number. In an attempt to define proteins and/or family of proteins that may either directly or indirectly influence DNA amplification potential through a mechanism of enhanced genomic instability, immobilized pH gradient-two-dimensional polyacrylamide gel electrophoresis (IPG 2-D PAGE) analysis of silver-stained nuclear cytoplasmic polypeptides concomitant with PALA resistance and CAD amplification was performed. Analysis of silver-stained polypeptides from 3 x LD50 PALA-selected CHO and HepG2 cells revealed no significant alterations in polypeptide expression. In CHO cells selected at 5 x and 7 x PALA LD50, and HepG2 cells selected at 5 x and 9 x PALA LD50, one subset of 4-8 polypeptides (pl: pI 7.2-7.6/36-38 kDa) were increased 2- to 3-fold in both 5 x and 7 x- and 5 x and 9 x LD50 PALA-selected CHO and HepG2, respectively, while five relatively neutral-to-basic, low M(r) polypeptides (p2: 18/7.30; p3: 16/7.00; p4: 14/7.00; p5: 14/7.40; and p6: 13.5/7.00) were markedly increased in CHO cells selected at 7 x LD50 PALA. In addition to these PALA-associated increases, four polypeptides (p7a: pI 6.50/40 kDa; p7b: 6.55/40; p7c: 6.60/40; and p7d: 6.65/40) were significantly increased in high-passage (p159) HepG2 cells undergoing spontaneous CAD gene amplification in the absence of PALA exposure. In CHO cells, polypeptides p7 a, b, d were increased while the expression of p7c (pI 6.60/40 kDa) was unaltered in 7 x LD50-treated CHO cells. Although neither the identity nor biological function of polypeptides 1-7 is known, a proposed mechanism involving interaction with certain growth regulatory proteins such as p53 for mediating genomic instability is given.
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PMID:Protein alterations associated with gene amplification in cultured human and rodent cells. 885 14