Purification and characterization of a plasminogen activator from mouse cells transformed by an oncogenic virus

Danø, Keld; Möller, Vanessa; Ossowski, Liliana; Nielsen, Lene

doi:10.1016/0005-2744(80)90110-2

Cited by 65 publications

(32 citation statements)

References 24 publications

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“…Extracts from a variety of murine tissues contain both u-PA and t-PA (19,41, and unpublished observations). Rabbit antibodies directed against murine u-PA completely inhibited its enzyme activity, without inhibiting t-PA (13).…”

Section: Resultsmentioning

confidence: 72%

“…All other materials were those described previously (2,3,10,13,16,19,41,42), or of the best commercially-available grade.…”

Section: Methodsmentioning

confidence: 99%

“…Controls were those recommended by Sternberger (48) and included (a) omission of either the first, second, or third layer of antiserum; (b) substitution of the primary antibodies by preimmune or nonimmune IgG (at concentrations of 5-50 ug/ml), or by control hyperimmune sera (antigastrin serum 4652, antisomatostatin serum R213/3) ; (c) absorption of the primary antibodies against varying concentrations (4 .2-63 KU/ml), of highly-purified murine prou-PA (3,41). The pro-u-PA used for antibody absorption showed one Coomassie Blue stainable band corresponding to M, 48 Kdaltons after SDS PAGE under reducing as well as nonreducing conditions (3) (detection limit for possible contaminating proteins -5%) .…”

Section: Methodsmentioning

confidence: 99%

“…We have reported the purification of murine u-PA (41), the development of rabbit antibodies against the purified enzyme (13), and the use of these antibodies for an immunocytochemical mapping ofthe distribution ofu-PA in the normal mouse (19). We now report their use for the immunocytochemical detection of u-PA in the invasivelygrowing, transplanted Lewis lung carcinoma.…”

mentioning

confidence: 99%

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Immunocytochemical localization of urokinase-type plasminogen activator in Lewis lung carcinoma.

Skriver¹,

Li²,

Kielberg³

et al. 1984

The Journal of Cell Biology

165

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The invasively growing and metastasizing Lewis lung carcinoma consistently contained urokinase-type plasminogen activator (u-PA) enzyme activity. When investigated immunocytochemically with antibodies against u-PA, different parts of individual tumors showed a pronounced heterogeneity in staining intensity. Strong staining was found in areas with invasive growth and degradation of surrounding normal tissue, while other areas were completely devoid of staining. Immunoreactivity occurred both with a perinuclear cytoplasmic localization in tumor cells and associated with apparently extracellular material. SDS PAGE of tumor extracts, under both reducing and nonreducing conditions, followed by immunoblotting, showed only one immunocytochemically stainable band with an electrophoretic mobility corresponding to that of purified proenzyme to u-PA, while no two-chain u-PA was detected. This indicates that the major part of the activator in Lewis lung carcinoma is present as one-chain pro-u-PA.

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Section: Resultsmentioning

confidence: 72%

“…All other materials were those described previously (2,3,10,13,16,19,41,42), or of the best commercially-available grade.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Immunocytochemical localization of urokinase-type plasminogen activator in Lewis lung carcinoma.

Skriver¹,

Li²,

Kielberg³

et al. 1984

The Journal of Cell Biology

165

View full text Add to dashboard Cite

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“…Conditioned serum-free culture fluid from the human fibrosarcoma cell line HT-1080 was prepared as reported for other cell lines [28], except that the cells were cultured in the presence of 10e6 M dexamethasone to increase inhibitor production [26]. Human pro-u-PA was prepared from the conditioned culture fluid of HT-1080 cells by affinity chromatography with a monoclonal antibody [29].…”

Section: Methodsmentioning

confidence: 99%

Plasminogen activators catalyse conversion of inhibitor from fibrosarcoma cells to an inactive form with a lower apparent molecular mass

Nielsen¹,

Andreasen²,

Grøndahl-Hansen³

et al. 1986

FEBS Letters

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Purified N 54 kDa plasminogen activator inhibitor from human fibrosarcoma cells was converted to an inactive form with slightly higher electrophoretic mobility by incubation with catalytic amounts of urokinasetype or tissue-type plasminogen activator. Serine proteinase inhibitors and a monoclonal antibody against urokinase-type plasminogen activator inhibited the conversion, indicating that it was caused by plasminogen activator-catalyzed proteolysis. These findings represent the first demonstration of a well-defined protein apart from plasminogen, constituting a substrate for plasminogen activators. PI~minogen activator Enzyme inhibitor (HT-1~8O~brosarcoma cell)

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Comparative study of plasminogen activator antigens in colonic carcinomas and adenomas

Suzumiya

Hasui

Kohga

et al. 1988

Intl Journal of Cancer

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The correlation between malignant transformation and increased PA synthesis or secretion has been examined in a variety of cell lines. To study the relationship between content and composition of PAs and colorectal neoplasms, we measured u-PA and t-PA antigen levels in normal mucosa, tubular adenoma, adenocarcinoma in adenoma, and adenocarcinoma, using a sensitive sandwich enzyme immunoassay. This assay was very sensitive and was not hindered by the presence of serine protease inhibitors. Both adenomas and carcinomas had higher u-PA antigen levels than normal mucosa. The u-PA antigen level of adenomas was lower than that of carcinomas. Antigen level of t-PA, however, was lower in both adenomas and carcinomas compared with that in normal mucosa, the values being lowest in carcinomas. Two cases of carcinoma in adenoma had PA contents similar to those of carcinomas. u-PA antigen level of adenomas with dysplastic epithelium was higher than that of adenomas without dysplastic epithelium. Therefore, the increase of u-PA content in adenomas could be a parameter of malignant changes in adenomas.

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Purification and characterization of a plasminogen activator from mouse cells transformed by an oncogenic virus

Cited by 65 publications

References 24 publications

Immunocytochemical localization of urokinase-type plasminogen activator in Lewis lung carcinoma.

Immunocytochemical localization of urokinase-type plasminogen activator in Lewis lung carcinoma.

Plasminogen activators catalyse conversion of inhibitor from fibrosarcoma cells to an inactive form with a lower apparent molecular mass

Comparative study of plasminogen activator antigens in colonic carcinomas and adenomas

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