Multicolor Imaging of Bifacial Activities of Estrogens
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...
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Published in | Methods in molecular biology (Clifton, N.J.) Vol. 1461; p. 153 |
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Main Authors | , |
Format | Journal Article |
Language | English |
Published |
United States
2016
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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. |
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AbstractList | 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. |
Author | Kim, Sung-Bae Umezawa, Yoshio |
Author_xml | – sequence: 1 givenname: Sung-Bae surname: Kim fullname: Kim, Sung-Bae email: kimu-sb@aist.go.jp organization: Research Institute for Environmental Management Technology, National Institute of Advanced Industrial Science and Technology (AIST), 16-1 Onogawa, Tsukuba, 305-8569, Japan. kimu-sb@aist.go.jp – sequence: 2 givenname: Yoshio surname: Umezawa fullname: Umezawa, Yoshio organization: Department of Chemistry, School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/27424902$$D View this record in MEDLINE/PubMed |
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Keywords | Estrogen receptor Multicolor imaging Bioluminescence Click beetle Luciferase Single-chain probe |
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SubjectTerms | Animals Cell Line Estrogens - genetics Estrogens - metabolism Gene Expression Gene Order Genetic Vectors - genetics Humans Ligands Luminescent Measurements - methods Molecular Imaging - methods Molecular Probes - chemistry Molecular Probes - genetics Protein Binding Protein Interaction Domains and Motifs Receptors, Estrogen - chemistry Receptors, Estrogen - genetics Receptors, Estrogen - metabolism |
Title | Multicolor Imaging of Bifacial Activities of Estrogens |
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