Unknown

Dataset Information

0

A dual-light reporter system to determine the efficiency of protein-protein interactions in mammalian cells.


ABSTRACT: Methods for determining protein-protein interactions in mammalian cells typically rely on single reporter functions and are susceptible to variations between samples particularly in regard to levels of transcription, processing and translation. A method has been developed for determining protein-protein interactions in mammalian cells, which bypasses these variables confounding single reporter assays. The approach utilizes two units of gene expression linked to reporter functions that are interposed by a deactivation-activation unit in such a way that the downstream expression unit is switched off. Hence upstream expression occurs regardless of protein-protein interaction, leading to the production of the upstream reporter. In the event of protein-protein interactions, the downstream expression unit is switched on leading to dual reporter read outs. Thus, the ratio of the two reporter activities provides a measure to determine the efficiency of protein-protein interactions. To access the system we screened a mutant of BMPR2 where the interaction between BMPR-II and LIMK is abrogated. BMPR-II is a type II receptor of the TGFbeta superfamily and plays a key role in the pathogenesis of familial pulmonary arterial hypertension. This system has potential for high-throughput screening of libraries (peptide, chemical, cDNA, etc.) to isolate agents that are capable of interfering with highly selective protein-protein interaction.

SUBMITTER: Nasim MT 

PROVIDER: S-EPMC1075926 | biostudies-literature | 2005

REPOSITORIES: biostudies-literature

altmetric image

Publications

A dual-light reporter system to determine the efficiency of protein-protein interactions in mammalian cells.

Nasim M T MT   Trembath R C RC  

Nucleic acids research 20050411 7


Methods for determining protein-protein interactions in mammalian cells typically rely on single reporter functions and are susceptible to variations between samples particularly in regard to levels of transcription, processing and translation. A method has been developed for determining protein-protein interactions in mammalian cells, which bypasses these variables confounding single reporter assays. The approach utilizes two units of gene expression linked to reporter functions that are interp  ...[more]

Similar Datasets

| S-EPMC7056976 | biostudies-literature
| S-EPMC4303541 | biostudies-literature
| S-EPMC10510171 | biostudies-literature
| S-EPMC8328604 | biostudies-literature
| S-EPMC5449462 | biostudies-literature
| S-EPMC6177421 | biostudies-literature
| S-EPMC8526717 | biostudies-literature
| S-EPMC7491154 | biostudies-literature
| S-EPMC4664949 | biostudies-literature
| S-EPMC3189979 | biostudies-literature