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:3.4.21.7 (plasmin)
9,023 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Tissue-type plasminogen activator (tPA) is a serine protease which cleaves plasminogen to its active form, plasmin. tPA plays a physiologic role in hemostasis, wound healing, and embryogenesis. Therapeutically, recombinant human tPA is used as a thrombolytic in myocardial infarction. Although production of therapeutic quantities of tPA in Chinese hamster ovary (CHO) cells transfected with the human gene for tPA is practical, production costs remain high. One important factor which determines the ultimate cost of tPA (or any other recombinant protein expressed in mammalian cells) is its production level on a per cell basis. We have used postembedding immunocytochemical staining with colloidal gold to study the subcellular localization of tPA in CHO cells expressing recombinant tPA (rCHO) in an effort to understand the factor(s) which might limit secretion. Staining for tPA was evaluated visually and by morphometric analysis and was specific and reproducible. Serially passaged rCHO showed no significant change in staining density over 31 serial passages. Staining density was greatest over dilated cisternae of the rough endoplasmic reticulum and nuclear envelope. Golgi stacks and large acid phosphatase-positive vacuoles (probably lysosomes) were also heavily stained. Staining of lysosomal vacuoles suggested that rCHO might be degrading nascent tPA. Incubation of rCHO with 125I-tPA showed that the cells were not internalizing tPA from the media. These results suggest that rCHO fail to secrete a portion of the tPA they synthesize and that it is degraded in lysosomes. This observation may have important implications on the choice of expression systems for efficient production of large quantities of recombinant proteins.
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PMID:Quantitative immunocytochemical staining for recombinant tissue-type plasminogen activator in transfected Chinese hamster ovary cells. 190 92

Evidence is presented for the involvement of a number of specific uterine- and conceptus-derived proteins in endometrial differentiation and conceptus or fetal development. These secretory proteins include mitogens (insulin-like growth factor-I and -II, epidermal growth factor, uterine luminal fluid mitogen), binding and transport proteins (uteroferrin, insulin-like growth factor and retinol binding proteins, respectively), protease inhibitors (antileukoproteinase, plasmin/trypsin inhibitor), and trophoblastic specific proteins. Using immunological reagents and specific complementary DNA (cDNA) probes, the tissue origins of several of these proteins have now been identified. In addition, the temporal regulation of messenger RNA (mRNA) production for a number of these proteins has been elucidated. The results suggest that although circulating and locally produced steroid hormones may be involved in regulating the synthetic abilities of these tissues during pregnancy, other, as yet undefined, factors may also mediate these activities. In this paper we present a review of the current knowledge pertaining to the identity, physiological regulation and potential functions of pig maternal and conceptus secretory proteins during pregnancy.
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PMID:Regulation of uterine and conceptus secretory activity in the pig. 219 44

The biosynthesis of distinct prostatic and lysosomal acid phosphatases is demonstrated using a human prostatic carcinoma cell line, PC-3SF12. The biosynthesis and maturation of the acid phosphatases was studied by metabolic labeling with radioactive leucine, specific immunoprecipitation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and fluorography. Of the tartrate-inhibitable acid phosphatase activity in PC-3SF12 cells, 60% is lysosomal and 10% is prostatic. The lysosomal-type acid phosphatase is synthesized as precursor with a molecular weight of 68,000, some of which is converted to higher-molecular-weight precursor polypeptides (Mr 71,000 and 77,000). The multiple forms of the precursors are due to differences in the carbohydrate chains on the enzyme because biosynthesis in the presence of tunicamycin eliminates the precursor multiplicity. The initial precursor (Mr 68,000) is processed to a mature polypeptide (Mr 49,000), via intermediates with molecular weights of 62,000 and 59,000. The mature polypeptide is degraded to smaller polypeptides with molecular weights of 30,000, 28,000, and 25,000. Precursor polypeptides of the lysosomal-type enzyme are secreted in the medium. Prostatic acid phosphatase is synthesized as a precursor with a molecular weight of 110,000, which is processed via several intermediates (Mr 99,000-93,000, 77,000, and 55,000) to a mature polypeptide with a molecular weight of 49,000. Particularly during cell homogenization, or lysis, the mature polypeptide is rapidly degraded to an immunoprecipitable polypeptide with a molecular weight of 20,000. None of these polypeptides is secreted in detectable amounts into the medium. Precursors and mature and smaller polypeptides are present in human prostate extract and seminal fluid. Proteolytic degradation of prostatic acid phosphatases in cells and tissues is probably catalyzed by a plasmin-like or related trypsin-like enzyme because degradation of the mature prostatic phosphatase polypeptide is completely prevented by addition of the plasmin inhibitor bovine pancreatic trypsin inhibitor. Prostatic- and lysosomal-type acid phosphatases are eventually stored at least in part in two different types of cell organelles. Testosterone does not influence the biosynthesis and secretion of either acid phosphatase in this cell line.
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PMID:Biosynthesis and processing of prostatic and lysosomal acid phosphatases in a prostate carcinoma cell line PC-3SF12. 294 40

We have compared the distribution of two of the major secreted proteins of the porcine uterus within the endometrium of ovariectomized pigs which had received hormone replacement therapy for 30 days. The proteins studied, plasmin/trypsin inhibitor (PI) and uteroferrin (Uf), an iron-containing acid phosphatase, were both secreted into the uterine lumen by ovariectomized gilts given progesterone (P4) or P4 and 17 beta-estradiol but not by animals given 17 beta-estradiol alone or corn oil. The two proteins were localized immunocytochemically within the endometrium using an immunoperoxidase procedure. The results confirmed that production of PI and Uf was P4-dependent and demonstrated that the primary site of synthesis of PI was the surface and upper glandular epithelium, while Uf synthesis was confined to the glandular epithelium. A similar localization of PI and of Uf was found in endometrial tissue from pigs at day 13 (late luteal phase) of the estrous cycle. These results suggest that the uterine epithelium of the pig is regionally differentiated with regard to the production of P4-induced proteins.
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PMID:Differential patterns of secretory protein localization within the pig uterine endometrium. 315 52

The uterus of the pig secretes large amounts of protein in response to progesterone. Estrogen alone has little effect but in combination with progesterone is synergistic at low doses and inhibitory at high doses. The responses of the uterus to progesterone require prolonged hormone treatment and are not immediate. The proteins secreted by the uterus of all species are believed to play some role in the nutritional and developmental support of the conceptuses, particularly during early pregnancy. Such a role is likely to be of greater importance in species such as the pig which possesses a noninvasive, diffuse-type of epitheliochorial placentation. A group of basic polypeptides dominates the uterine secretions of the pig. The best characterized is uteroferrin, a purple colored, iron-containing acid phosphatase which transports iron across the placenta. Three polypeptides which are found associated noncovalently with uteroferrin have been shown to be antigenically closely related to each other and to have arisen from a single precursor polypeptide. Their function is unknown. A family of plasmin/trypsin inhibitors which show sequence homology with bovine pancreatic trypsin inhibitor (aprotinin) has been well characterized and appears to control intrauterine proteolytic events initiated by the conceptuses. Several other proteins secreted in response to progesterone remain to be characterized and functionally defined.
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PMID:Hormonal control and function of secretory proteins. 345 17

In Exp. 1, administration of 5 mg oestradiol valerate i.m. to pregnant gilts on Days 9 or 9 and 10 advanced the uterine secretion of calcium, protein, and acid phosphatase as demonstrated by levels recovered in the uterine flushings of females unilaterally hysterectomized on Day 11. Upon removal of the remaining uterine horn on Day 12, protein and acid phosphatase increased while Ca2+ decreased in oestradiol-treated gilts as did PGF. In contrast, a 4-fold increase in recoverable Ca2+ occurred from Days 11 to 12 in control gilts. Recoverable oestradiol-17 beta was increased in all 3 groups on Day 12 and plasmin inhibitor concentration increased in oestradiol-treated gilts. Two-dimensional PAGE demonstrated the appearance of a group of very acidic polypeptides in oestradiol-treated gilts. Blastocysts recovered from the second uterine horn had undergone elongation to the filamentous morphology in all 3 groups. In Exp. 2, oestradiol valerate was administered to pregnant gilts on Day 9 or Days 9 and 10 followed by total hysterectomy on Day 16. No differences in recoverable Ca2+ or protein were found, but acid phosphatase was decreased by 75% after oestradiol treatment. Recoverable oestradiol was decreased in oestradiol-treated gilts while PGF and plasmin inhibitor concentrations were unaffected. Compared with the control gilts, blastocysts recovered from oestradiol-treated gilts were fragmented and degenerating on Day 16. PAGE demonstrated greatly intensified staining of the group of acidic polypeptides in oestradiol-treated gilts. These results indicate that oestradiol treatment on Day 9 of pregnancy advances uterine secretory response, but that blastocyst elongation can occur in this uterine environment and in the presence of declining intraluminal Ca2+ levels.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Development and survival of pig blastocysts after oestrogen administration on day 9 or days 9 and 10 of pregnancy. 359 51

The porcine uterus secretes a group of basic, low molecular weight protease inhibitors under the influence of progesterone, but not estrogen. One of these inhibitors (Mr approximately 14,500) which inhibits trypsin, plasmin, and chymotrypsin, but not other proteases tested, has been purified 10- to 15-fold from uterine secretions of pseudopregnant pigs using Sephadex G-100 chromatography, CM-cellulose ion exchange chromatography, and Sephadex G-50 or Bio-Gel P-10 chromatography. The inhibitor which is relatively heat- and pH-stable forms a 1:1 molar complex with trypsin which is not dissociated in sodium dodecyl sulfate except by boiling. Chymotrypsin appears to bind at the same site on the inhibitor as trypsin. The inhibitor is high in half-cysteine residues and basic amino acids, and appears not to be a glycoprotein. Antiserum has been raised against the purified inhibitor in rabbits and used to test its distribution in pigs using the immunoperoxidase-staining technique on tissue sections. The inhibitor is associated only with the glandular and surface epithelium of the uterus. Endometrial explants from pseudopregnant animals, cultured in presence of L-[3H]leucine, release the inhibitor in radioactive form indicating that it is a uterine product. The antiserum against the inhibitor cross-reacts with at least three other, basic, low molecular weights plasmin/trypsin inhibitors in porcine uterine secretions, suggesting that a family of isoinhibitors exists which may constitute up to 15% of the protein in porcine uterine secretions. The inhibitor(s) appears to coat and to be taken up by the trophoectoderm cells of the elongating blastocyst during pregnancy. It is suggested that the inhibitors may serve to protect the uterus from proteases released by the porcine trophoblast or to prevent degradation of essential macromolecules, such as uteroferrin, which have to be taken up by the conceptus.
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PMID:Purification and properties of a progesterone-induced plasmin/trypsin inhibitor from uterine secretions of pigs and its immunocytochemical localization in the pregnant uterus. 621 38

Using a modified model of Masugi's nephritis of rats, various enzymatic activities in urine, serum and renal tissue (glomeruli or cortex) were determined at appropriate intervals after the administration of anti-kidney serum and compared with the urinary protein content and the kidney weight. In the urine, alkaline phosphatase (Al-Phosase), acid phosphatase (Ac-Phosase) and N-acetyl-beta-glucosaminidase (NA-beta-Gase) activities remarkably increased after the induction of nephritis, reached their peaks on the 10th day and reverted to almost the normal levels on the 30th day. The patterns of time course of these enzymatic activities were similar to patterns seen in the urinary protein content and the kidney weight. In the serum, the Al-Phosase activity decreased slightly, while NA-beta-Gase activity increased slightly. The Ac-Phosase activity in serum remained at normal levels during the experimental periods. In the glomeruli, the bound activities of these three enzymes decreased with nephritis, showing a negative correlation with results in the urine. On the other hand, fibrinolytic activities in the urine (plasmin-like enzyme) and renal cortex (plasminogen activator) also paralleled the urinary protein content and the kidney weight in the course of the disease. These results suggest that the Al-Phosase, Ac-Phosase and NA-beta-Gase excreted into urine in cases of nephritis may be mostly derived from damaged renal cells and one part of Al-Phosase may also come from the plasma. Moreover, the increase of plasmin-like enzyme in urine is considered to be due to the increase of plasminogen activator in the renal cortex. Thus, the determination of these enzymatic activities in the urine should be useful for evaluating effects of drugs for the treatment of nephritis.
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PMID:Pharmacological studies on experimental nephritic rats (9). Changes in activities of urinary enzymes in the modified type of Masugi's nephritis and their sources. 720 60

The porcine uterus synthesizes a proteinase inhibitor (M(r) 14,000) under the influence of progesterone that is relatively specific for plasmin and trypsin, but that also has weak affinity for chymotrypsin. Several isoforms of this uterine plasmin/trypsin inhibitor were purified by a procedure whose final two steps involved affinity chromatography on immobilized chymotrypsin and cation exchange chromatography. Amino-terminal sequencing showed that at least three of the isoforms were closely related. An oligonucleotide probe based on the protein sequence was used to identify a cDNA that contained an open reading frame coding for a mature protein (M(r) 10,295) of 93 amino acids. The inhibitor had a well defined, but unique, Kunitz domain of 64 residues at its amino terminus that shared 67% sequence identity to bovine pancreatic trypsin inhibitor. Its P1 residue was arginine rather than lysine. Northern analysis showed the presence of a single mRNA species (700 bases) that in adult female pigs appeared to be confined to the uterus. During pregnancy, UPTI mRNA expression was high until Day 30 and decreased significantly thereafter. By contrast, uteroferrin mRNA reached maximal concentrations in late pregnancy. These data are consistent with an earlier hypothesis that the inhibitor serves to neutralize the activities of one or more serine proteinases generated by the proliferating trophoblast during the formation of the noninvasive placenta of the pig.
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PMID:Purification, characterization, and cDNA cloning of a Kunitz-type proteinase inhibitor secreted by the porcine uterus. 792 61

Objectives were to examine the effects of a single dose (4 mg) of estradiol-17 beta (E2) on blastocyst development around the period of elongation. Proestrus gilts were induced to ovulate with 750 IU of hCG and were mated before ovulation (normal mating, 24 to 32 h post-hCG) or after ovulation had begun (delayed mating, 43 h post-hCG). This difference in time of mating has been demonstrated to result in approximately a 7-h difference in time of blastocyst elongation. Normally and delay-mated gilts were ovariohysterectomized at 278 h post-hCG or injected with E2 or vehicle (corn oil) at 278 h and then ovariohysterectomized at 290 h post-hCG (five or six gilts per group). Blastocyst size was measured and concentrations of E2, retinol, uteroferrin, insulin-like growth factor-I (IGF-I), uterine plasmin/trypsin inhibitor (UPTI) and protein in uterine flushings were quantified. Blastocyst size and components of uterine flushings did not differ (P > 0.05) between normally and delay-mated gilts at 278 h post-hCG. However, at 290 h post-hCG, normally mated gilts had larger (P < 0.01) blastocysts (small spheres to filamentous) and their flushings tended to contain less (P < 0.07) amounts of retinol than those of delay-mated gilts whose blastocysts ranged from small spheres to ovoidals. Normally mated gilts receiving E2 at 278 h had smaller (P < 0.01) blastocysts and less (P < 0.05) amounts of retinol at 290 h post-hCG than gilts receiving vehicle. Conversely, delay-mated gilts treated with E2 or vehicle did not differ (P > 0.05) in blastocyst size and amounts of components of uterine flushings at 290 h post-hCG. Normally mated gilts treated with vehicle had litters in the process of elongating at 290 h post-hCG. Mean blastocyst size (P < 0.001) and amounts of components of uterine flushings (except for IGF-I) in these gilts were greater (P < 0.05, UPTI = 0.06) than in normally mated gilts at 278 h post-hCG, whose blastocysts were spherical. Among gilts not treated with E2 (278 h and 290 h pooled), mean blastocyst size was positively correlated (P < 0.05) with amounts of retinol, E2, uteroferrin and total protein. Results indicated that a single dose of E2 given before elongation altered blastocyst development depending on how close blastocysts were to onset of elongation at the time of E2 treatment.
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PMID:Short-term effects of exogenous estradiol-17 beta on blastocyst development during the period of elongation in swine. 923 12


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