Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.4.21.4 (trypsin)
42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Pancreatitis was induced in 11 miniature pigs by infusing a bile salt-trypsin solution into the pancreatic duct. Seven animals served as sham-operated controls. Serum ionized calcium, total calcium, albumin, total protein, inorganic phosphorus, urea nitrogen, magnesium, insulin, glucagon, and hematocrit were determined every six to 12 h over a period of one week in both test and control animals. We observed significant decreases in ionized and total calcium, modest decreases in albumin, and significant increases in the inorganic phosphorus, urea nitrogen, and hematocrit in the pancreatitic pigs. The latter two findings were consistent with early acute hypovolemia. Glucagon and insulin appeared to play no role in the hypocalcemia. Glucagon concentrations increased to the same degree in both test and control animals, probably as a result of the stress of being handled and operated on. The highest concentrations of inorganic phosphorus and the lowest concentrations of both ionized and total calcium were seen 18 h after the induction of pancreatitis in the test animals. These findings suggest that parathyrin (parathormone) was not being secreted in adequate amounts, or that the target organs were unresponsive to parathyrin.
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PMID:Biochemical changes in a porcine model of acute pancreatitis. 65 76

Low salt extracts from homogenates of bovine cardiac muscle contain two protease inhibitors, one specific for the calcium-activated protease from this tissue and the other for trypsin and chymotrypsin, but no other serine proteases, including plasmin, thrombin, and subtilisin. The former, which can be separated from the protease by chromatography on DEAE-cellulose, is a protein with a molecular weight of 270,000. Its action is not based on the sequestering of calcium, and it is present in large excess over the amount of calcium-activated protease in this tissue. The trypsin inhibitor, which has a molecular weight of 70,000, is estimated to be present at approximately 300 microgram/g, wet weight, of tissue. The identification of inhibitors such as these in the cytoplasm may explain why nonlysosomal proteolytic activity has been thought to be insignificant in the overall turnover of intracellular protein and suggests that a re-evaluation of this possibility is necessary.
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PMID:Identification of two protease inhibitors from bovine cardiac muscle. 68 25

Native spectrin has trypsin-susceptible sites spaced at a constant molecular weight interval. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of spectrin treated with trypsin at low salt concentrations shows a ladder of fragments spaced at approximately 8,000-dalton intervals, from the intact Band 1 (240,000 daltons) and Band 2 (220,000 daltons) down to about 150,000 daltons. The five largest fragments were identified as products of Band 2 using tryptic 125I-peptide mapping of protein from gel slices. Endogenously incorporated [32P]phosphate is absent from the largest fragment, indicating that all phosphorylation sites on spectrin are within 8,000 daltons of a terminal of Band 2. Mapping of both [14C]carboxyamidomethylated cysteine-containing tryptic peptides and 125I-peptides reveals extensive sequence homology between the spectrin subunits. Further, only somewhat over half of the distinct spots expected from the cysteine content are found in both Band 1 and Band 2 peptides. These and the tryptic susceptibility results are interpretable as evidence for a repeating structure in spectrin.
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PMID:Structural studies on human spectrin. Comparison of subunits and fragmentation of native spectrin. 76 4

Sporocysts of Isospora endocallimici, a parasite of marmosets, were exposed to minimal essentials medium (MEM) or a trypsin-bile salt solution (TBS) and then fixed and prepared for transmission electron microscopy. Excystation occurred in TBS but not MEM. The sporocyst wall has 2 layers, a thin outer layer (15 to 110 nm thick) and a thick inner layer (65 to 180 nm thick), which is composed of 4 separate curved plates. The outer layer consists of 1 to 3 membranes interspersed with lipid droplets. In the inner layer, a thin layer of material connects the peripheral margins of 2 apposing plates. Immediately beneath this layer, a thin strip of material is interposed between the 2 apposing plates. Ultrastructural changes preparatory to excystation occur primarily in the inner layer of the sporocyst wall. The TBS acts upon the site of apposition between 2 plates causing the interposed strip to swell and separate from the margin of each plate which leads to collapse of the sporocyst. As the sporocyst collapses, the margins of each curved plate curl inward toward the center of the sporocyst.
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PMID:Ultrastructure of the sporocyst wall during excystation of Isospora endocallimici. 82 16

Pancreatic and biliary secretion and gastric emptying rates of a liquid test meal (LTM) were determined in normal persons, in patients with subtotal gastrectomy with gastroduodenostomy (STG-BI) or with gastrojejunostomy (STG-BII), and in patients with truncal vagotomy and pyloroplasty (V&P). In all operated persons, rapid gastric emptying diluted intraluminal contents, with consequent abnormally low concentrations of trypsin and bile salts initially, a pattern that was not corrected by addition of intravenous hormones to the meal stimulus. Trypsin output in V&P's after the LTM was significantly depressed to 40% of normal, but was normal in the STG groups. The delay in reaching normal values for trypsin and bile salt concentrations, was more marked in STG-BII owing to sequestration of secretions in the afferent loop. The low luminal concentrations of digestive secretions for the first 60 to 80 min after a LTM are therefore attributable to rapid gastric emptying in all operated groups, and in V&P to a depressed pancreatic enzyme response also. In STG-BII, afferent loop sequestration exaggerates the delay in attainment of normal intraluminal concentrations. The combined disturbance in STG-BII produces greater abnormalities than seen in STG-BI.
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PMID:Gastric emptying of liquid meals and pancreatic and biliary secretion after subtotal gastrectomy or truncal vagotomy and pyloroplasty in man. 83 May 68

Leopard shark triacylglycerol lipase has been characterized as a crude pancreatic preparation. The enzyme demonstrated an absolute requirement for trihydroxy bile salts for activity with natural bile salts of the shark giving a 4-fold greater stimulation of activity than pure sodium taurocholate. Bile salts also protected the enzyme from apparent inactivation by p-chloromercuribenzoate and trypsin treatment. The shark lipase demonstrated a temperature optimum of 36 degrees C and was rapidly inactivated at 50 degrees C even in the presence of bile salts. Divalent metal ions were required for activity with Ca2+ providing the greatest stimulation. At 22 degrees C, pH 8.5 and in the presence of natural bile salts, the apparent V was about 0.6 mumol fatty acid released/min per mg protein. The shark enzyme hydrolyzed over 90% of the fatty acids from trioleovylglycerol and methyl esters of pancreatic lipase-resistant fatty acids were hydrolyzed at the same rate as typical fatty acid methyl esters. Hydrolysis of triacylglycerol proceeded about ten-times faster than wax ester hydrolysis. The kinetic properties of the leopard shark enzyme were compared to other bile salt-dependent lipolytic enzymes. Pancreatic lipase activity was not detected.
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PMID:Partial characterization of the bile salt-dependent triacylglycerol lipase from the leopard shark pancreas. 83 61

It was previously shown that calf uterus cytosol contains a Ca2+-activated receptor transforming factor (RTF) which irreversibly converts the larger molecular states of estrogen receptor (5.3 to 8.6 S, depending on ionic strength) into a smaller, salt-stable form (4.5 S, independent of ionic strength). We now describe a method for rapid and reliable separation of precursor and RTF-transformed receptor forms, which takes advantage of a difference in isoelectric point between the two: the more acidic precursor (isoelectric point, 6.2) is still retained by DEAE-cellulose under conditions (0.12 M KCl, pH 8.3) which produce release from cellulose of the less acidic transformed form (isoelectric point, 6.6 to 6.8). Based on this method of separation, RTF activity can be assayed easily and we could thus progress in the purification and physical and functional characterization of this factor, RTF has been purified about 100-fold. Molecular properties, as assayed by methods suited to partially purified preparations, are as follows: sedimentation coefficient, 6.4 S; Stokes radius, 45 A; molecular weight, 115,000; isoelectric point, 4.9. The Michaelis constant, expressed as moles/liter of estradiol binding sites, is 1.25 X 10(8), at pH 7.5 and 4 degrees, pH 8.5 is optimum for activity. RTF attacks native casein (Km, 1.25 X 10(-5) mol/liter at pH 7.5 and 22 degrees) but not hemoglobin, ovalbumin, or albumin. N-Benzoylarginine methyl ester is a competitive inhibitor of RTF-induced receptor transformation, while L-leucylglycylglycine and N-benzoyltyrosinamide are not. RTF activity is protected by -SH compounds. RTF activity is Ca2+-dependent. Ca2+ starts an activation-inactivation cycle of RTF, with permanent loss of transforming activity which proceeds at a particularly fast rate in the absence of substrate. Mg2+ is inactive, while Sr2+ and Mn2+ may in part substitute for Ca2+. RTF is present in both endometrium and myometrium. RTF is not a lysosomal hydrolase, as shown by its alkaline pH optimum (8.5) and exclusive location in cytosol, nor is it trypsin or a protease of the trypsin group. Also, it is distinct from known proteases of human uterus. The functional significance of this Ca2+-activated protease of cytosol with alkaline pH optimum and high affinity for the larger native form of receptor is still unknown.
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PMID:Estrogen binding proteins of calf uterus. Molecular and functional characterization of the receptor transforming factor: A Ca2+-activated protease. 83 20

The dexamethasone-binding receptor protein in rat liver cytosol has a Stokes radius of 61 A and a sedimentation coefficient of 4.0 S. In contrast, cell nuclei labelled with [3H]dexamethasone in vivo or in vitro (reconstitution experiments with [3H]dexamethasone-labelled cytosol and isolated unlabelled nuclei) contain a high-salt-extractable dexamethasone-receptor complex with a Stokes radius of 30-36 A and a sedimentation coefficient of 3.2 S. Exposure of liver homogenate or 1000 X g homogenate supernatant to low ionic strength during preparation of cytosol resulted in conversion of the 61 A to a 36 A complex very similar to the intranuclear form of dexamethasone receptor. 61 leads to 36 A complex-converting activity was present in both the 100 X g-10 000 X g sediment of liver homogenate, from which it could be extracted by hypotonic media, and in the liver cell nuclei, from which it could be extracted by hypertonic media. Mild digestion of the 61 A dexamethasone-receptor complex with trypsin also gave rise to a complex with a Stokes radius of 36 A. Reconstitution experiments with isolated liver cell nuclei indicated that both the 61 A and 36 A dexamethasone-receptor complexes were taken up by the nuclei; reextraction of the nuclei incubated with the 61 A complex revealed that this form had been converted to the 30-36 A complex. Further digestion of the 61 and 36 A [3H]dexamethasone-receptor complexes with hypotonic extract of the 1000 X g-10 000 X g sediment of liver homogenate or with trypsin resulted in formation of a third complex with Stokes radius of 19 A and a sedimentation coefficient of 2.5 S. The approximate molecular weights of the 61, 36 and 19 A dexamethasone-receptor complexes were calculated as 102 000, 46 000 and 19 000, respectively, and the frictional ratios of the molecules as 1.84, 1.38 and 1.00, respectively. It is concluded that the nuclear 30-36 A dexamethasone-receptor complex is formed from the cytosol 61 A complex by proteolytic digestion and that this latter protein contains at least two sites with a relatively high sensitivity to protelytic cleavage.
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PMID:Formation and characteristics of hepatic dexamethasone-receptor complexes of different molecular weight. 87 74

Procedures were developed for isolating highly purified cytoplasmic granules of basophilic leukocytes from guinea pig peripheral blood. The methods involved disruption of cells in 0.34 M sucrose followed by a series of membrane filtrations and fractionation on sucrose density gradients. These preparations, up to 95% pure basophil granules by electron microscopy, contained a mixture of neutral esterases-proteases including caseinolytic activity; both trypsin- and chymotrypsin-like serine hydrolases were identified by means of appropriate inhibitors. Localization of at least one such activity to the basophil granule was confirmed by a cytochemical method; this activity was absent in contaminating lymphocytes and eosinophils. By contrast, several lysosomal enzymes, lactic dehydrogenase, and plasminogen activator activity, present in cell homogenates, were absent from purified granules. The granule matrix of guinea pig basophils, unlike the cytoplasmic granules of other granulocytes or mast cells, was little altered by high or low salt concentration but was disrupted into insoluble fragments by 0.01 N HCl and by Triton X-100. Granules were solubilized by papain and by urea-SDS but enzyme activity was destroyed. Triton X-100 incubation with freeze-thawing proved to be the optimal method for extracting esterase activities. Esterase activities were not released from basophils under conditions of anaphylactic degranulation that liberated the great majority of basophil granule histamine.
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PMID:Isolation of the cytoplasmic granules of guinea pig basophilic leukocytes: identification of esterase and protease activities. 87 25

A bile salt mixture and pure sodium taurocholate were each shown to increase the esterolytic activity of trypsin in aqueous solution and in intestinal juice. rho-Toluene-sulphonyl-L-arginine methyl ester (TAME) was used as a substrate, and both a spectrophotometric and a potentiometric assay system were used. The maximal potentiation of the esterolytic activity of trypsin by bile salts was about 1.6 to 2.2 times the activity without bile salts (depending on the assay conditions and whether the trypsin was in aqueous solution or intestinal juice). The proteolytic activity of trypsin was decreased by the addition of bile salts. It seemed likely, therefore, that the potentiating effect of bile salts on trypsin esterolytic activity is primarily on the substrate (TAME) rather than trypsin itself. It was thought that TAME might be taken up into bile salt micelles and thus be more readily hydrolysed by trypsin, but we were unable to substantiate this hypothesis. The precision of the trypsin esterolytic assay was better when bile salts were not added. If however bile salts were to be used routinely in the trypsin assay, it would be useful to ensure that the concentration of calcium, included as activator, is sufficiently low to prevent the formation of a precipitate. This precipitate is probably a complex of calcium and bile salts.
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PMID:The effect of bile salts on the esterolytic assay of trypsin. 91 20


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