title: SH2 Domain-Based FRET Biosensor for Measuring BCR-ABL Activity in Living CML Cells.
abstract: Fluorescent proteins (FPs) displaying distinct spectra have shed their light on a wide range of biological functions. Moreover, sophisticated biosensors engineered to contain single or multiple FPs, including Förster resonance energy transfer (FRET)-based biosensors, spatiotemporally reveal the molecular mechanisms underlying a variety of pathophysiological processes. However, their usefulness for applied life sciences has yet to be fully explored. Recently, our research group has begun to expand the potential of FPs from basic biological research to the clinic. Here, we describe a method to evaluate the responsiveness of leukemia cells from patients to tyrosine kinase inhibitors using a biosensor based on FP technology and the principle of FRET. Upon phosphorylation of the tyrosine residue of the biosensor, binding of the SH2 domain to phosphotyrosine induces conformational change of the biosensor and brings the donor and acceptor FPs into close proximity. Therefore, kinase activity and response to kinase inhibitors can be monitored by an increase and a decrease in FRET efficiency, respectively. As in basic research, this biosensor resolves hitherto arduous tasks and may provide innovative technological advances in clinical laboratory examinations. State-of-the-art detection devices that enable such innovation are also introduced.
authors.affiliation: Department of Cell Physiology, Hokkaido University Graduate School of Medicine, N15W7, Kita-ku, Sapporo, 060-8638, Japan.
authors.forename: Mari
authors.initials: M
authors.lastname: Fujioka
authors.affiliation: Tamano Technology Center, Research and Development Headquarters, Mitsui Engineering & Shipbuilding Co., Ltd., 3-16-1, Tamahara, Tamano, 706-0014, Japan.
authors.forename: Yumi
authors.initials: Y
authors.lastname: Asano
authors.affiliation: Tamano Technology Center, Research and Development Headquarters, Mitsui Engineering & Shipbuilding Co., Ltd., 3-16-1, Tamahara, Tamano, 706-0014, Japan.
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Nakada
authors.affiliation: Department of Cell Physiology, Hokkaido University Graduate School of Medicine, N15W7, Kita-ku, Sapporo, 060-8638, Japan. yohba@med.hokudai.ac.jp.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Ohba
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D001854:Bone Marrow Cells / Q000187:drug effects / Q000378:metabolism; D005434:Flow Cytometry; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation / Q000379:methods*; D016044:Fusion Proteins, bcr-abl / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects; D017930:Genes, Reporter; D006801:Humans; D007091:Image Processing, Computer-Assisted; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010766:Phosphorylation; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D011993:Recombinant Fusion Proteins; D012984:Software; D014162:Transfection; D014407:Tumor Cells, Cultured; D018909:src Homology Domains
chemical_list: D011993:Recombinant Fusion Proteins; D016044:Fusion Proteins, bcr-abl
keywords: BCR-ABL; Chronic myeloid leukemia (CML); CrkL; Fluorescence cytometry (FCM); Fluorescence lifetime; Fluorescence microscopy; Fluorescent protein (FP); Förster resonance energy transfer (FRET); Molecular target drug; Src Homology 2 (SH2) domain