Chapter title |
Multicolor Imaging of Bifacial Activities of Estrogens.
|
---|---|
Chapter number | 12 |
Book title |
Bioluminescence
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3813-1_12 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3811-7, 978-1-4939-3813-1
|
Authors |
Sung-Bae Kim, Yoshio Umezawa |
Editors |
Sung Bae Kim |
Abstract |
The present protocol introduces multicolor imaging of bifacial activities of an estrogen. For the multicolor imaging, the authors fabricated two single-chain probes emitting green or red bioluminescence (named Simer-G and -R, respectively) from click beetle luciferase (CBLuc) green and red: Simer-R consists of the ligand binding domain of estrogen receptor (ER LBD) and the Src homology-2 (SH2) domain of Src, which are sandwiched between split-CBLuc red (CBLuc-R). On the other hand, Simer-G emitting red light consists of the ER LBD and a common consensus sequence of coactivators (LXXLL motif), which are inserted between split-CBLuc green (CBLuc-G). This probe set creates fingerprinting spectra from the characteristic green and red bioluminescence in response to agonistic and antagonistic activities of a ligand of interest. The present protocol further provides a unique methodology to calculate characteristic estrogenicity scores of various ligands from the spectra. |
You are seeing a free-to-access but limited selection of the activity Altmetric has collected about this research output.
Click here to find out more.