Synthesis in vitro of urokinase-like material using polyA(+)RNA from human kidney and from cultured human embryonic kidney cells

Bollen, Alex; Glineur, Cécile; Herzog, Albert

doi:10.1016/s0006-291x(80)80155-0

Cited by 8 publications

(3 citation statements)

References 9 publications

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“…Recently, the induction of a large amount of the urokinase was demonstrated in DNA-repair-deficient cells that had undergone UV irradiation (5,21). Human urokinase is synthesized as a single-chain proenzyme in the kidney and probably is secreted in urine (4,29,34). The amino acid sequences and structure of this enzyme have been well studied (8,9,25,33,39).…”

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confidence: 99%

A monoclonal antibody against human urokinase: Characterization of the epitope and its localization in human kidney.

Nakamura

Takahashi

Naora

et al. 1984

Cell Struct. Funct.

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ABSTRACT. Monoclonal antibodies against human plasminogen activator urokinase have been produced. A G62 hybridoma-producing antibody (IgG) was purified on a DEAE-cellulose column, and it proved useful for the measurement, identification and purification of antigens that had approximate molecular weights of 55-and 33-Kdaltons.For immunochemical measurements and purification, a competitive enzymelinked immunosorbent assay (ELISA) and affinity chromatography using antibody-immobilized Sepharose 4B were developed. The ELISA has sensitivity to 20 p mole antigen molecules. The binding capacity of the antigen on the affinity column was evaluated on SDS-polyacrylamide slab gels as well as by fibrin autography and ELISA. Results showed that there was quantitative purification with no loss of enzyme activity in the one-step procedure.Western blotting and affinity binding showed antigenic bands with apparent molecular weights of 55-and 33-Kdaltons. Because the 55-Kdalton form contains 33-and 22-Kdalton components connected by a disulfide bond, the epitope domain is present on the 33-Kdalton chain.Using this antibody, we examined human kidney sections by direct immunofluorescence to locate the antigen. It was found in epithelial cells of convoluted segments, in glomerulus cells and in capillary endothelial cells, evidence that renal tubular cells synthesize the antigen which then is secreted in urine.

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confidence: 99%

A monoclonal antibody against human urokinase: Characterization of the epitope and its localization in human kidney.

Nakamura

Takahashi

Naora

et al. 1984

Cell Struct. Funct.

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“…3) and embryonic kidney cells (data not shown). The discrepant results obtained by other authors (24,25) may be due to insufficiently specific antisera. In addition, because no other bands of higher molecular weight are precipitated by anti-urokinase antibodies, the higher molecular weight forms of urokinase (22,23) do not seem to represent biosynthetic precursors of urokinase.…”

Section: Discussionmentioning

confidence: 74%

“…These authors also report that this mRNA is very abundant in embryonic kidney cells, up to 10% of the total poly(A)+ RNA. Another group has reported that proteins of -47,000 and 20,000 daltons and none of higher mass, can be immunoprecipitated by an anti-urokinase antiserum from the proteins synthesized in an in vitro cellfree system directed by embryonic or adult human kidney mRNA (25). The latter authors estimated the amount of urokinase mRNA to be -10% of the total kidney poly(A)+ RNA.…”

Section: Discussionmentioning

confidence: 99%

Monoclonal antibodies to human urokinase identify the single-chain pro-urokinase precursor.

Salerno¹,

Verde²,

Nolli³

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

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Monoclonal antibodies have been obtained that recognize either the A or B chain of human urinary urokinase. These antibodies identify human urokinase-producing cells and the product of urokinase mRNA. Anti-urokinase monoclonal antibodies precipitate an =w54,000-dalton protein synthesized in vitro in a rabbit reticulocyte cell-free system. This pro-urokinase appears to be the precursor of both A and B chains of human urinary urokinase. Urokinase mRNA in human kidney constitutes only 0.1% or less of total poly(A)+ RNA.

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Expression in Escherichia coli of urokinase antigenic determinants

Bollen¹,

Glineur²,

Delcuve³

et al. 1981

Biochemical and Biophysical Research Communications

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Synthesis in vitro of urokinase-like material using polyA(+)RNA from human kidney and from cultured human embryonic kidney cells

Cited by 8 publications

References 9 publications

A monoclonal antibody against human urokinase: Characterization of the epitope and its localization in human kidney.

A monoclonal antibody against human urokinase: Characterization of the epitope and its localization in human kidney.

Monoclonal antibodies to human urokinase identify the single-chain pro-urokinase precursor.

Expression in Escherichia coli of urokinase antigenic determinants

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