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)

A specific, sensitive and accurate radioimmunoassay (RIA) method for the measurement of prostaglandin A1 (PGA1) in either human whole blood or plasma is described. Whole blood is immediately lysed with distilled water containing tritiated indicator. When plasma is assayed, the blood samples are handled at 4 C and rapidly centrifuged. The lysate or plasma is adjusted to pH 5 with buffer and quickly extracted with 5% methanol in dichloromethane. The whole blood or plasma extract is then purified by Sephadex LH20 chromatography using the system methanol: methylene chloride (5:95) which separates the major groups of PGA, PGE and PGF. The RIA is then performed using an antiserum generated in rabbits from PGA1 coupled to bovine thyroglobulin. The antibody is highly specific, possessing very low cross reactivity to other prostaglandins (PGA2, PGE, PGB and PGF). Activated florisil or ammonium sulfate can be used to separate bound from free prostaglandin. This whole blood or plasma method yields blank values of only 2 +/- 2 pg per sample with a between assay precision determined by duplicate analysis of 8% and interassay precision of 3%. The mean whole blood PGA1 concentration in 27 subjects in 2.5 +/- 1.6 (SD) ng per 100 ml. No significant sex difference in PGA1 levels was noted and values were similar whether measured in whole blood or cooled plasma rapidly prepared and extracted. These values of PGA1 are much lower than those RIA values reported by others for "PGA" using antibodies with lower specificities.
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PMID:A radioimmunoassay for prostaglandin A1 in human peripheral blood. 115 43

1. Two pepsins, designated Pepsin I and Pepsin II, were isolated and partially characterized from the stomach of the adult stage salmon Oncorhynchus keta. This stage is developed in a marine environment. 2. One pepsin, designated Pepsin II, was isolated from the stomach of the juvenile stage salmon Oncorhynchus keta. This stage is developed in an estuarine environment. 3. The enzymes were partially purified by ammonium sulfate precipitation, ion exchange chromatography and gel filtration. 4. Pepsins I and II from adults and Pepsin II from juvenile showed proteolytic activity on acid-denatured hemoglobin with a pH optimum of 3. 5. The mol. wt determined by gel filtration on Sephadex G-100 of Pepsin I from juvenile species was found to be 32,000 whereas a value of 27,000 was determined for Pepsin II from juvenile and adult fish. 6. In contrast with Pepsin II, Pepsin I was activated by NaCl. It is suggested that the appearance of NaCl-activated pepsin would represent and adaptive response of the organism to the change from a low to a high salinity environment.
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PMID:Partial purification of pepsins from adult and juvenile salmon fish Oncorhynchus keta. Effect of NaCl on proteolytic activities. 311 85

In a radioassay for Vasoactive Intestinal Peptide (VIP)-binding, eight out of 33 plasma samples from healthy human subjects exhibited specific binding ranging from 2.6% to 46.7% of total [125 I]VIP. This binding was competitively displaced by unlabeled VIP. The structurally homologous peptides, Peptide Histidine Isoleucine (PHI) and secretin, were, respectively, 72-fold and 413-fold less potent than VIP in displacing bound [125 I]VIP, whereas the unrelated peptides, neurotensin, eledoisin, bombesin and metenkephalin, were without effect on the binding. The antibody nature of the VIP-binding factor was suggested by its precipitation with ammonium sulfate, attenuation after absorption with Staphylococcus aureus preparations, precipitation with antisera against human IgG and IgM, and coelution with standard IgG and IgM on anion-exchange and high-performance gel-filtration columns. Pepsin treatment of purified IgG fraction yielded a VIP-binding species with apparent molecular weight of 108 +/- 13 kDa that was precipitated by antiserum against the F(ab)2 fragment of the IgG molecule. These results demonstrate the existence in some human plasmas of an autoantibody that binds VIP.
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PMID:Autoantibody to vasoactive intestinal peptide in human circulation. 383 70

1. The abomasum of the milk-fed calf has been examined using an adaptation of the Serial Test Meal method devised by Hunt & Spurrell (1951). The emptying process, acid secretion and pepsin secretion were studied.2. Using serial test meals of simple solutions instilled into the abomasum via a cannula, our investigation leaves no doubt that the osmolarity of the abomasal contents significantly modifies the rate of abomasal emptying.3. Hypotonic and isotonic solutions of sodium chloride and sodium bicarbonate increase abomasal emptying but bicarbonate is most effective.4. Increasing the concentration of solutes in the abomasal contents slows abomasal emptying. Sodium chloride, sodium bicarbonate, ammonium chloride and urea do not delay abomasal emptying until hypertonic concentrations are attained. Hypotonic solutions of potassium chloride, calcium chloride, glucose, lactose, hydrochloric acid and acetic acid delay abomasal emptying.5. The results obtained in the calf show that the abomasum is under restraint probably from duodenal receptors as is the simple stomach (Hunt & Knox, 1968) and that an osmoreceptor as postulated by Hunt (1956) is an important factor in this mechanism.6. Acid secretion is inhibited when hypertonic solutions are instilled into the abomasum.7. Pepsin secretion is not affected by simple solutions in the abomasum.8. Gastric function in the milk-fed calf appears to be controlled by mechanisms essentially similar to those already demonstrated in the simple stomach.
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PMID:The effect of some molecules and ions on gastric function in the milk-fed calf. 456 11

Camel (Camelus dromedarius) pepsins were precipitated from the extract of the fundic gastric mucosa by ammonium sulfate between 35 and 80% saturation. DEAE--cellulose chromatography of this fraction produced two isoenzymes, I and II, which were further purified to homogeneity on sephadex G-100. Their Km with N-acetyl-L-phenylalanyl-diiodotyrosine were 0.10 and 0.90 mM, and their molecular weights which were determined on sodium dodecylsulfate gel electrophoresis exhibited 35,500 and 34,700, respectively. The two pepsins were essentially free of carbohydrates, but contained 0.3 and 1.0 mol of organic phosphate per 1 mol of protein, respectively. The apparent mobilities of the phospho- and dephosphoforms of each pepsin were indifferent in polyacrylamide gel electrophoresis at pH 8.9. The N-terminal residues of pepsins I and II were found to be alanine and leucine, respectively. Pepsin I was inactivated at a faster rate than that of pepsin II at pH 8 and 0 degrees C, and at pH 7.5 and 37 degrees C; but both were denatured under these conditions. The properties of these enzymes are compared with those of other mammalian and avian pepsins.
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PMID:Isolation and characterization of camel pepsins. 614 Oct 27

A collagenous protein could be precipitated by (NH4)2SO4 from the culture medium of a murine teratocarcinoma-derived cell line (Ko, C.Y., Johnson, L.D. and Priest, R.E. (1979) Biochim. Biophys. Acta 581, 252-259). Further purification of this protein was achieved by combining DEAE-cellulose chromatography with either CM-cellulose or molecular sieve chromatography. The collagenous polypeptides had subunit molecular weights of 160 000, if determined by molecular sieve chromatography, or 190 000, if determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and they are not linked by disulphide bridges. Amino acid composition of this collagen is similar to that of a murine type IV collagen isolated from EHS sarcoma (Timpl et al. (1978) Eur. J. Biochem. 84, 43-52). The most prominent peptides resulting from cleavage of the protein by CNBr had estimated molecular weights of 25 000, 23 000, 11 700 and 9400. Pepsin treatment of this collagen under non-denaturing conditions produced three major fragments having molecular weights of 70 000, 45 000 and 43 000. We conclude that the collagen secreted by the murine teratocarcinoma-derived cell culture is a type IV basement membrane collagen. Therefore, this culture system should provide a continuous source of type IV collagen, which may be used to study the interaction of this collagen with other basement membrane components.
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PMID:Purification and characterization of a collagenous protein secreted by a murine teratocarcinoma-derived cell line. 710 99

Isoelectric focusing revealed three enolase isoforms in pig brain, which were designated as alphaalpha-(pI = 6.5), alphagamma- (pI = 5.6), and gammagamma-enolase (pI = 5.2). The pI of purified gammagamma-enolase was also 5.2. The gammagamma-enolase isoform of enolase was purified from pig brain by a purification protocol involving heating to 55 degrees C for 3 min, acetone precipitation, ammonium sulfate precipitation (40%-80%), DEAE Sephadex ion-exchange chromatography (pH 6.2), and Sephadex G200 gel filtration. The final specific activity was 82 units/mg protein. As with other vertebrate enolases, gammagamma-enolase from pig proved to be a dimer with a native mass of 85 kDa and a subunit mass of 45 kDa. The pH optimum for the reaction in the glycolytic direction is 7.2. The Km values for 2-PGA, PEP, and Mg2+ were determined to be 0.05, 0.25, and 0.50 mM, respectively, similar to Km values of other vertebrate enolases. The amino acid composition of pig gammagamma-enolase, as determined by amino acid analysis, shows strong similarity to the compositions of gammagamma-enolases from rat, human, and mouse, as determined from their amino acid sequences. Despite the differences seen with some residues, and considering the ways that the compositions were obtained, it is assumed that pig gammagamma-enolase is more similar than the composition data would indicate. Moreover, it is likely that the sequences of pig gammagamma-enolase and the other gammagamma-enolases are almost identical. Li+ proved to be a noncompetitive inhibitor with either 2-PGA or Mg2+ as the variable substrate. This enolase crystallized in the monoclinic space group P2, or P2(1). An Rsymm <5% was obtained for data between 50 and 3.65 A, but was a disappointing 30% for data between 3.65 and 3.10 A, indicating crystal disorder.
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PMID:Purification and properties of gammagamma-enolase from pig brain. 1007 35

Bacillus licheniformis ATCC 9945a is one of the bacterial strains that produce gamma-poly(glutamic acid) (gamma-PGA). The use of carbohydrate medium components for gamma-PGA production was explored. Cells were grown in shake flasks or in controlled pH fermentors using medium formulations that contain different carbon sources. During the cultivations, aliquots were removed to monitor cell growth, carbon utilization, polymer production, and polymer molecular weight. Glucose was a better carbon source than glycerol for cell growth. Furthermore, glucose was utilized at a faster rate than glycerol, citrate, or glutamate. However, by using mixtures of glucose and glycerol in medium formulations, the efficiency of gamma-PGA production increased. For example, by increasing the glycerol in medium formulations from 0 to 40 g/L, the gamma-PGA broth concentration after 96 h increased from 5.7 to 20.5 g/L. Considering that glycerol utilization was low for the glucose/glycerol mixtures studied, it was unclear as to the mechanism by which glycerol leads to enhanced product formation. Cell growth and concomitant gamma-PGA production (12 g/L) at pH 6.5 was possible using glucose as a carbon source if trace amounts (0.5 g/L each) of citrate and glutamate were present in the medium. We suggested that citrate and glutamate were useful in preventing salt precipitation from the medium. In addition, glutamate may be preferred relative to ammonium chloride as a nitrogen source. The conversion of glucose to gamma-PGA by the strain ATCC 9945a was believed to occur by glycolysis of glucose to acetyl-CoA and tricarboxylic acid (TCA) cycle intermediates that were then metabolized via the TCA cycle to form alpha-ketoglutarate, which is a direct glutamate precursor.
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PMID:Effects of glucose and glycerol on gamma-poly(glutamic acid) formation by Bacillus licheniformis ATCC 9945a. 1009 19

Cytosolic pyruvate kinase (ATP: Pyruvate phosphotransferase, EC 2.7.1.40; PKc) was purified to apparent homogeneity with about 22% recovery from developing seeds of Brassica campestris using (NH4)2SO4 fractionation, DEAE-cellulose chromatography, gel filtration through Sepharose-CL-6B and affinity chromatography through reactive Blue Sepharose-CL-6B. The purified enzyme with molecular mass of about 214 kDa was a heterotetramer with subunit molecular mass of 55 and 57 kDa. The enzyme showed maximum activity at pH 6.8 and absolute requirement for a divalent (Mg2+) and a monovalent (K+) cation for activity. Typical Michaelis-Menten kinetics was obtained for both the substrates with Km values of 0.10 and 0.11 mM for PEP and ADP, respectively. The enzyme could also use UDP or GDP as alternative nucleotides, but with lower Vmax and lesser affinities. The enzyme was inhibited by glutamate, glutamine, fumarate, citrate, isocitrate, oxalate, 2-PGA, ATP, UTP and GTP and activated by glucose-6-phosphate, fructose-1,6-bisphosphate and Pi, suggesting its regulation mainly by TCA cycle intermediates and the cellular need for carbon skeletons for amino acid biosynthesis. ATP inhibition was of competitive type with respect to PEP and non-competitive with respect to ADP. Similarly, oxalate inhibition was also of competitive type with respect to PEP and non-competitive with respect to ADP. Initial velocity and product inhibition studies except for pyruvate inhibition were consistent for a compulsory-ordered tri-bi mechanism.
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PMID:Purification and characterization of cytosolic pyruvate kinase from developing seeds of Brassica campestris L. 1098 13

Poly(gamma-D-glutamic acid) (PGA)-producing strains of Bacillus species were investigated to determine their ability to contribute to reducing the amount of ammonium nitrogen in liquid manures and their ability to convert some of the ammonium into this polyamino acid as a transient depot for nitrogen. Organisms that do these things should help solve the serious environmental problems which are caused by the use of large amounts of liquid manure resulting from intensified agriculture; these problems are mainly due to the high content of ammonium nitrogen. Bacillus licheniformis ATCC 9945 and Bacillus subtilis were able to grow in liquid manure and to produce PGA in the presence of sodium gluconate. On artificial liquid manure these two strains were able to produce 0.85 and 0.79 g of PGA per liter, respectively. Under conditions that are found in intensified farming situations the ammonia content was reduced within 48 h from 1.3 to 0.75 g/liter. One mutant of B. subtilis 1551 impaired in the catabolism of PGA was obtained after nitrosoguanidine mutagenesis. This mutant produced PGA at a final concentration of 4.8 g/liter, whereas the wild type produced only 3.7 g/liter.
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PMID:Cultivation of bacteria producing polyamino acids with liquid manure as carbon and nitrogen source. 1115 24


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