Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0272170 (SDS)
 50,377 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Our previous study showed that an epitope defined by a monoclonal antibody against human urokinase is located on the 33-Kdalton catalytic domain of the enzyme (Nakamura, M. et al., Cell Struct Funct., 9, 167-179, 1984). The epitope structure was further determined and characterized on one-dimensional SDS-polyacrylamide slab gel maps of CNBr-cleaved polypeptide fragments as well as on their Western blots. A single homogeneous polypeptide with an approximate molecular weight of 3.4-Kdaltons was found to be antigenic. The monoclonal antibody exhibited a stronger inhibition of the enzyme activity than the polyclonal antibodies tested, and cross-reacted with a 65-Kdalton tissue-type plasminogen activator present in Detroit 562 cells. From these results and data made up with the help of a computer comparison of known sequences of urokinase and a tissue-type plasminogen activator, we concluded that the epitope is Cys-Gln-Gly-Asp-Ser-Gly-Gly-Pro-Leu-Val-Cys and contains a catalytically active residue, serine.
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PMID:A monoclonal antibody against human urokinase: the epitope structure and sequence homology with a human tissue-type plasminogen activator. 241 11

Mesangial cells in culture change shape and become less adhesive in response to cAMP elevation (e.g., treatment with isoproterenol plus isobutylmethylxanthine (IM). Inhibitors of serine proteases inhibit cellular shape change in response to IM. To further examine the role of cell surface proteases in shape change, adhesion plaque proteins (i.e., preparations of ventral membranes and extracellular matrix) were separated in SDS-polyacrylamide gels containing gelatin with and without plasminogen. Four discrete zones of lysis were evident in plasminogen gels (indicative of activation of plasminogen) from control adhesion plaques: one inconspicuous zone with a Mr approximately 150 kD, another at approximately 115 kD, and a doublet at approximately 35-32 kD. Another diffuse zone of lysis centered around Mr approximately 70 kD and contained a defined band of approximately 56 kD. Adhesion plaques contained most of the plasminogen activators (PA). 5 min after IM treatment, the Mr approximately 150- and approximately 115-kD PA were increased in activity. Vasopressin (VP), which prevented shape change and adhesion loss when added along with IM, inhibited the increase in these PA. Preincubation with monoclonal or polyclonal antibodies to urokinase-type plasminogen activator (uPA) totally inhibited the IM-inducible shape change and adhesion loss. Activation of plasminogen throughout the gels revealed multiple protease resistant bands that markedly increased with IM treatment (maximal at 45 min). These may represent focal control mechanisms. uPA thus may mediate focal proteolysis, which results in shape change and decreased adhesion.
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PMID:Urokinase-dependent adhesion loss and shape change after cyclic adenosine monophosphate elevation in cultured rat mesangial cells. 246 65

The amino terminal fragment (ATF) of urokinase-type plasminogen activator (uPA) is a degradation product comprising the entire growth factor-like and kringle domains. It has been previously shown that ATF is able to bind to the u-PA receptor through the growth factor-like domain and that the anti u-PA monoclonal antibody 5B4 (Mab 5B4) binds to ATF preventing u-PA receptor binding. To localize more precisely the epitope recognized by Mab 5B4, ATF was subfragmented by controlled enzymatic proteolysis with V8 protease. Three subfragments of 4,000 Mr (F-4k), 11,000 Mr (F-11k) and 12,000 Mr (F-12k) were purified from the reaction mixture and characterized. SDS-PAGE under reducing and non-reducing conditions, N-terminal aminoacid sequence analysis and C-terminal aminoacid analysis of each fragment indicate that F-4k and F-11k correspond to intact growth factor-like domain and kringle domain (residues 4-43 and 44-135 respectively) while F-12k corresponds to the kringle domain cleaved in the first loop at the glu52-gly53 bond. By Western blot and competitive binding experiments we show that Mab 5B4 recognizes an epitope located on the kringle domain of u-PA and that the binding is strongly reduced when the kringle contains an additional cleavage in its first loop. Since the receptor binding site of u-PA has been previously shown to be located on the growth factor-like domain, Mab 5B4 inhibits the binding of uPA to its cellular receptor likely by steric hindrance. Besides the proven utility in epitope localization of anti u-PA monoclonal antibodies, these u-PA fragments may represent powerful tools for studies of structure-function relationship of u-PA.
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PMID:Epitope mapping of the anti-urokinase monoclonal antibody 5B4 by isolated domains of urokinase. 248 Jun 54

The plasminogen activator (PA) activity produced by Syrian hamster embryo (SHE) cells in different stages of neoplastic conversion was analysed. PA activity was characterized immunologically and by SDS-PAGE. Normal SHE cells had a very low PA activity. Although activity of either the tissue type of PA (t-PA) or the urokinase type (u-PA) or both were found to be increased in most immortal or transformed SHE cells, there was no correlation between enhanced production of a particular PA type and the development of the immortal or transformed phenotype. However, within a group of cell lines clonally derived from a culture of immortal cells, a positive correlation was found between extracellular t-PA, but not u-PA, activity and cellular growth rate. For the Syrian hamster PA species, crossreacting with anti-human u-PA, a mol. wt of 39 kd was observed. For the Syrian hamster PA species, crossreacting with anti-human t-PA, multiple species were found with mol. wts of 98, 72 and 59 kd respectively. Evidence was obtained that the 72-kd species represents the intact enzyme, the 59-kd species a partial digestion product thereof and the 98 kd species, which often appears as a doublet, a complex of either of these species with an inhibitor, likely to be secreted by the same cells. Finally, our data suggest a novel mechanism for the enhancement of t-PA activity of transformed cells, namely by a decrease in the effective extracellular amount of putative inhibitor.
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PMID:Enhanced plasminogen activator production of Syrian hamster embryo cells transformed by chemicals or the c-Ha-ras oncogene: type of plasminogen activators involved and their contribution to the transformed phenotype. 250 Feb 66

Plasminogen activator inhibitor (PAI) was purified in active form from porcine platelets under nondenaturing conditions. The purified inhibitor (Mr 47,000) reacts with tissue-type plasminogen activator (t-PA), urokinase (UK), and activated protein C (APC) to yield both SDS-stable complexes and a modified PAI of slightly reduced molecular weight. The second-order rate constants for the inhibition of t-PA and UK by PAI are 3.5 X 10(7) and 3.4 X 10(7) M-1 s-1, respectively. Activated protein C reacts with PAI with a second-order rate constant of 1.1 X 10(4) M-1 s-1. This rate is not accelerated by protein S, phospholipid, and calcium, or heparin. It is concluded that (1) PAI can function as both inhibitor and substrate of its target proteases, (2) if APC promotes fibrinolysis via inactivation of PAI, then APC must be present in concentrations several orders of magnitude greater than t-PA, or the interaction of APC and PAI must be accelerated by presently unknown mechanisms, and (3) in the absence of heparin, platelet PAI is the most rapid inhibitor of APC yet described.
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PMID:Platelet plasminogen activator inhibitor: purification and characterization of interaction with plasminogen activators and activated protein C. 250 42

Testicular cells from human biopsy specimens were cultured and plasminogen activator secreted into the medium was investigated. Follicle stimulating hormone stimulated plasminogen activator secretion. SDS-polyacrylamide gel electrophoresis followed by zymography showed that the molecular weight of plasminogen activator produced by human testicular cells is the same as that of human urokinase. Antigenicity was also found to be of the urokinase type but not of the tissue type.
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PMID:Secretion of plasminogen activator in response to follicle-stimulating hormone in culture medium of human testicular cells from biopsy specimens. 250 63

Immature mice were injected subcutaneously with 5 IU PMSG for 2 days to stimulate follicle development, which was followed by administration of 5 IU hCG to induce ovulation. The ovaries were removed at various periovulatory stages for preparing ovarian homogenates, granulosa cells and cumulus-oocyte complexes. The activity of plasminogen activator in the samples, separated by SDS-PAGE, were determined by fibrin-overlay technique. The results show that 15% of the gonadotropin-treated animals were ovulated 8h after hCG administration, about 6-8h earlier than that occurred in rat. Moreover, both tPA, and uPA activity were stimulated following PMSG treatment in ovarian homogenates and granulosa cells. Subsequent hCG injection further increased the two types of PA activity in a time-dependent manner, reaching maximum 4-8h after hCG treatment, and declined following ovulation. Greater uPA activity (70%) in the cultured mouse granulosa cells was found. It is, therefore, suggested that both tPA and uPA may be involved in the regulation of ovulation in mouse. The cumulus-oocyte complexes contained mainly tPA, which activity showed a time-dependent increase and reached a maximum between 12-24h after hCG treatment. Since cumulus-oocyte complexes collected from oviducts post ovulation still retain a considerable amount of tPA, the enzyme in the complexes may also play a role in the process of cumulus dispersion, oocyte transportation and implantation.
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PMID:[Plasminogen activator activity in mouse ovaries during periovulatory period]. 250 47

Human chromosomal DNA encoding single-chain urokinase-type Plasminogen Activator (scu-PA, or pro-urokinase) was inserted in an expression plasmid and transfected in human A431, mouse LB6 and CHO cells. LB6 cells were also transfected with a Bovine Papilloma Virus derivative containing the scu-PA gene. Human scu-PA was purified from cell supernatants of recombinant clones and characterized for structure and function. All recombinant scu-PAs are undistinguishable from human urine-derived scu-PA for peptide backbone, but possess a higher sugar content, as revealed by SDS-PAGE analysis after digestion with glycopeptidase F. This difference is partly due to an increased sialic acid content, as shown by analysis of neuraminidase-treated scu-PAs. No difference was found, however, among recombinant and natural scu-PAs in the kinetics of conversion into two-chain active forms (tcu-PAs) by human plasmin, and in the KM and kcat values of tcu-PA activity on the chromogenic substrate S-2444 and on human plasminogen. Also, recombinant and non-recombinant tcu-PAs displayed similar dose-response curves for binding to the endothelial inhibitor PAI-1. In conclusion, the glycosylation pattern of u-PA does not affect its interaction with the plasma proteins directly involved in its fibrinolytic function.
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PMID:The differential glycosylation of human pro-urokinase from various recombinant mammalian cell lines does not affect activity and binding to PAI-1. 251 81

SDS-polyacrylamide gel electrophoresis followed by enzymography of a human testicular cell culture medium demonstrated that the molecular weight of plasminogen activator (PA) secreted was similar to that of human urokinase and it was also confirmed that follicle-stimulating hormone (FSH) stimulated PA secretion. To further investigate the correlation between spermatogenic impairment and secretory potential of PA in human testicular cells in vivo, the maximum increment in PA activity as a result of treatment with ovine FSH was measured for two groups of patients with different degrees of spermatogenic impairment. The mean value of maximum increment in PA activity in one group with severely impaired testes was remarkably lower than that in another group with slightly impaired testes. Protein biosynthesis in human testicular cells dependent on FSH appears to be impaired with the progression of spermatogenic impairment.
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PMID:Plasminogen activator activity of testicular cells of subfertile men and FSH in vitro. 251 18

Guerin epithelioma, a highly metastatic tumour implanted to Wistar rats contains two inhibitors of fibrinolysis which can be detected with the use of zymographic techniques. The first one--with Mr about 48000 forms SDS-stable complex with urokinase. The second--with Mr about 7000 inhibits fibrinolytic and amidolytic activity of plasmin.
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PMID:Fibrinolytic inhibitors from the experimental rat epithelioma. 252 18


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