Protein-protein interactions (PPIs) are the basis of cell communication and the backbones of signal pathways, which can cause various diseases when abnormal. Therefore, identifying and regulating PPIs can unlock the pathophysiological mechanism of disease and make important contributions to the development of potential new drugs. The mammalian two-hybrid system (M2H) is a variant of the yeast two-hybrid system (Y2H). Similar to Y2H, the principle of M2H relies on the fusion of the cDNAs of interest with both the DNA-binding domain (DNA-BD) and the DNA-activation domain (DNA-AD), named bait and prey plasmid, respectively These plasmids are then introduced into appropriate host cells together with the reporter plasmid.
According to statistics, about 50% of the interactions described in PubMed are determined by Y2H. However, Y2H has an inevitable limitation, that is, even if there is no interaction during screening, false-positives and false-negatives and automatic activation of reporter genes are often recorded. The development of M2H is to better overcome these drawbacks and provide the environment for the bona fide posttranslational modification and localization of most eukaryotic proteins. Since the technology was first developed, several M2H applications have been developed, especially focusing on identifying deregulated signaling pathways in diseases.
Figure 1. Conventional mammalian two-hybrid assay (Patrício, D.; Fardilha, M. 2020)
Creative Proteomics has recruited many experienced molecular biology experts and technicians to manage our mammalian two-hybrid platform. On our platform, customers can more efficiently study mammalian protein interactions in an environment which is that is more similar to that in vivo. In addition to identifying potential interactions, the platform can also identify residues/domains involved in protein-protein interactions, as well as small molecules that affect protein-protein interactions.
In order to help customers complete projects with higher quality, we have also developed many adaptations to conventional M2H, such as mammalian PPI traps (MAPPIT), reverse MAPPIT, mammalian membrane two-hybrid assay, etc.
Customers can choose different technology platforms according to project requirements, or contact us directly for consultation, and our expert team will provide you with customized experimental procedures.
Creative Proteomics is an international biotechnology company dedicated to research in molecular interactions and other related fields. The mammalian two-hybrid platform we constructed has the characteristics of high quality and efficiency, and the data obtained can be directly used for paper publication. Our one-stop service aims to save customers time and money.