We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Utilization of an Intracellular Calcium Mobilization Assay for the Screening of Transduced FK506-Binding Proteins.
- Authors
Cha, Hyun Ju; Lee, Hyunjin; Yeo, Eun Ji; Yeo, Hyeon Ji; Choi, Yeon Joo; Sohn, Eun Jeong; Kim, Dae Won; Park, Soo Jung; Lee, Sung Ho; Lee, Sunghou; Choi, Soo Young
- Abstract
FK506-binding proteins (FKBPs) belong to the immunophilin family and are linked to various disease states, including the inflammatory response. The inhibition of cytokine and chemokine expression in addition to positive effects of FKBPs on corneal inflammation in animal models suggests that they may be used for ophthalmic delivery in the treatment of dry eye disease. To pass the effective barriers protecting eye tissues, testing the transduction domains of FKBPs is essential. However, monitoring their transduction efficiencies is not a simple task. The quantitative measurement of FKBP interactions was performed using a cell model with a specific G protein-coupled receptor, as FKBPs had been known to act at the inositol 1,4,5-trisphosphate receptor (IP3R) leading to the inhibition of intracellular calcium mobilization. Because of its luminescence amplitude and stability, human urotensin II receptor was expressed in aequorin parental cells to measure the action of selected FKBPs. This luminescence-based functional assay platform exhibited a high signal-to-background ratio of more than 100 and a Z' factor at 0.6204. As expected, changes in the sequence of the transduction domain affected the function of the FKBPs. The intracellular calcium mobilization assay with selected FKBPs represented a robust and reliable platform to screen initial candidates. Although the precise nature of the control that FKBPs exert on the IP3R is uncertain, this approach can be used to develop innovative anti-inflammatory treatments for dry eye disease by optimizing protein transduction domain sequences.
- Subjects
INTRACELLULAR calcium; DRY eye syndromes; ANIMAL models of inflammation; PROTEIN domains; G protein coupled receptors; PROTEINS
- Publication
Assay & Drug Development Technologies, 2021, Vol 19, Issue 7, p442
- ISSN
1540-658X
- Publication type
Article
- DOI
10.1089/adt.2021.065