Cloning and expression of the gene for the major excreted protein of transformed mouse fibroblasts. A secreted lysosomal protease regulated by transformation.

Troen, Bruce R.; Ascherman, Dana P.; Atlas, Daphine; Gottesman, Michael M.

doi:10.1016/s0021-9258(19)57386-0

Cited by 57 publications

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References 33 publications

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“…This fragment was inserted into the SacII-XhoI site of the eucaryotic expression vector pCOl (provided by D. Lowy) to yield pHaMDR1, with the human MDR1 cDNA sequences inserted between the two long terminal repeats of Harvey murine sarcoma virus. pcosMEP5 is a genomic clone of the mouse MEP gene in the cosmid vector pSV13 (19,30). pHu16 is a cDNA clone of the human MEP sequences in the Okayama-Berg expression vector (8a, 21).…”

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confidence: 99%

Use of a Cloned Multidrug Resistance Gene for Coamplification and Overproduction of Major Excreted Protein, a Transformation-Regulated Secreted Acid Protease

Kane

Troen

Gal

et al. 1988

Molecular and Cellular Biology

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Malignantly transformed mouse fibroblasts synthesize and secrete large amounts of major excreted protein (MEP), a 39,000-dalton precursor to an acid protease (cathepsin L). To evaluate the possible role of this protease in the transformed phenotype, we transfected cloned genes for mouse or human MEP into mouse NIH 3T3 cells with an expression vector for the dominant, selectable human multidrug resistance (MDR1) gene. The cotransfected MEP sequences were efficiently coamplified and transcribed during stepwise selection for multidrug resistance in colchicine. The transfected NIH 3T3 cell lines containing amplified MEP sequences synthesized as much MEP as did Kirsten sarcoma virus-transformed NIH 3T3 cells. The MEP synthesized by cells transfected with the cloned mouse and human MEP genes was also secreted. Elevated synthesis and secretion of MEP by NIH 3T3 cells did not change the nontransformed phenotype of these cells.

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confidence: 99%

Use of a Cloned Multidrug Resistance Gene for Coamplification and Overproduction of Major Excreted Protein, a Transformation-Regulated Secreted Acid Protease

Kane

Troen

Gal

et al. 1988

Molecular and Cellular Biology

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Cysteinyl Proteinases and Their Selective Inactivation

Shaw

1990

Advances in Enzymology - And Related Areas of Molecular Biology

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A Novel Photoaffinity‐Based Probe for Selective Detection of Cathepsin L Active Form

et al. 2012

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Detecting the active forms of proteases by using activity-based probes in complex proteomes has become an intensively investigated field of research over the past years because many pathogenic conditions involve alterations in protease activities. The detection of lysosomal cysteine proteases, the cathepsins, has mostly relied on the use of probes that incorporate reactive electrophilic moieties to modify a cysteine in the active site covalently. Here we report the first example of an activity-based probe that targets the cathepsins and incorporates a photoactivatable benzophenone group for covalent labelling. When tested on a set of five cathepsins (B, K, L, S and V), this probe selectively labelled the active site of cathepsin L. Furthermore, when tested on crude cell extracts, the probe specifically detected cathepsin L quantities as low as a few picomoles. This study suggests that photoaffinity labelling is a promising approach for developing highly selective and useful cathepsin L probes. In particular, this probe might allow the detection of small amounts of the secreted active cathepsin L form in the cellular microenvironment in vitro and ex vivo.

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Cloning and expression of the gene for the major excreted protein of transformed mouse fibroblasts. A secreted lysosomal protease regulated by transformation.

Cited by 57 publications

References 33 publications

Use of a Cloned Multidrug Resistance Gene for Coamplification and Overproduction of Major Excreted Protein, a Transformation-Regulated Secreted Acid Protease

Use of a Cloned Multidrug Resistance Gene for Coamplification and Overproduction of Major Excreted Protein, a Transformation-Regulated Secreted Acid Protease

Cysteinyl Proteinases and Their Selective Inactivation

A Novel Photoaffinity‐Based Probe for Selective Detection of Cathepsin L Active Form

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