Studying Chemoattractant Signal Transduction Dynamics in Dictyostelium by BRET.
Understanding the dynamics of chemoattractant signaling is key to our understanding of the mechanisms underlying the directed migration of cells, including that of neutrophils to sites of infections and of cancer cells during metastasis. A model frequently used for deciphering chemoattractant signal transduction is the social amoeba Dictyostelium discoideum. However, the methods available to quantitatively measure chemotactic signaling are limited. Here, we describe a protocol to quantitatively study chemoattractant signal transduction in Dictyostelium by monitoring protein-protein interactions and conformational changes using Bioluminescence Resonance Energy Transfer (BRET).
Department of Chemistry and Biochemistry, University of Arizona, 1041 E. Lowell Street, Tucson, 85721-0088, AZ, USA.
A F M Tariqul
AF
Islam
Department of Chemistry and Biochemistry, University of Arizona, 1041 E. Lowell Street, Tucson, 85721-0088, AZ, USA.
Branden M
BM
Stepanski
Department of Chemistry and Biochemistry, University of Arizona, 1041 E. Lowell Street, Tucson, 85721-0088, AZ, USA. pcharest@email.arizona.edu.
Pascale G
PG
Charest
D060807:Bioluminescence Resonance Energy Transfer Techniques; D002465:Cell Movement; D002630:Chemotactic Factors; D002633:Chemotaxis; D000242:Cyclic AMP / Q000378:metabolism; D004023:Dictyostelium / Q000502:physiology*; D017930:Genes, Reporter; D020962:Heterotrimeric GTP-Binding Proteins / Q000378:metabolism; D011485:Protein Binding; D025941:Protein Interaction Mapping; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D015203:Reproducibility of Results; D015398:Signal Transduction; D014170:Transformation, Genetic
D002630:Chemotactic Factors; D043562:Receptors, G-Protein-Coupled; D011993:Recombinant Fusion Proteins; D000242:Cyclic AMP; D020962:Heterotrimeric GTP-Binding Proteins
Bioluminescence resonance energy transfer; Chemotaxis; Dictyostelium discoideum; Directed cell migration; GPCR; Heterotrimeric G proteins; Protein–protein interaction; Signaling dynamics