| Chapter title |
The Use of Two-Photon FRET-FLIM to Study Protein Interactions During Nuclear Envelope Fusion In Vivo and In Vitro.
|
|---|---|
| Chapter number | 7 |
| Book title |
The Nuclear Envelope
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3530-7_7 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3528-4, 978-1-4939-3530-7
|
| Authors |
Richard D. Byrne, Banafshé Larijani, Dominic L. Poccia |
| Editors |
Sue Shackleton, Philippe Collas, Eric C. Schirmer |
| Abstract |
FRET-FLIM techniques have wide application in the study of protein and protein-lipid interactions in cells. We have pioneered an imaging platform for accurate detection of functional states of proteins and their interactions in fixed cells. This platform, two-site-amplified Förster resonance energy transfer (a-FRET), allows greater signal generation while retaining minimal noise thus enabling application of fluorescence lifetime imaging microscopy (FLIM) to be routinely deployed in different types of cells and tissue. We have used the method described here, time-resolved FRET monitored by two-photon FLIM, to demonstrate the direct interaction of Phospholipase Cγ (PLCγ) by Src Family Kinase 1 (SFK1) during nuclear envelope formation and during male and female pronuclear membrane fusion in fertilized sea urchin eggs. We describe here a generic method that can be applied to monitor any proteins of interest. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 7 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Librarian | 1 | 14% |
| Other | 1 | 14% |
| Professor | 1 | 14% |
| Student > Ph. D. Student | 1 | 14% |
| Researcher | 1 | 14% |
| Other | 1 | 14% |
| Unknown | 1 | 14% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 4 | 57% |
| Biochemistry, Genetics and Molecular Biology | 1 | 14% |
| Social Sciences | 1 | 14% |
| Chemistry | 1 | 14% |