RRC ID 48811
Author Inoue H, Sakaue T, Ozawa T, Higashiyama S.
Title Spatiotemporal visualization of proHB-EGF ectodomain shedding in living cells.
Journal J Biochem
Abstract Heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF) is a member of the EGF family, each of which is produced as a type I transmembrane precursor. The juxtamembrane domain of proHB-EGF, a precursor of HB-EGF, is cleaved by a disintegrin and metalloproteases. HB-EGF is released into the extracellular space and strongly activates EGF receptor. The relevance of better understanding proHB-EGF shedding relates to the importance of the process in the proliferation, differentiation and survival of various types of cells. Shedding of proHB-EGF is normally evaluated using an alkaline phosphatase-tagged proHB-EGF assay or a western blotting assay that involves multiple cells, which makes it difficult to observe spatiotemporal differences in the activities of the individual cells. In this study, we developed a fluorescent proHB-EGF-based metalloprotease biosensor, named Fluhemb, to visualize spatiotemporal regulation of proHB-EGF shedding in individual cells using a simple method that measures changes in fluorescence ratios. Fluhemb might be very useful for detecting the activity of proHB-EGF shedding in various types of cells under different conditions in vitro and in vivo.
Volume 154(1)
Pages 67-76
Published 2013-7-1
DOI 10.1093/jb/mvt030
PII mvt030
PMID 23598347
MeSH Animals Biosensing Techniques / methods Cell Line Cell Membrane / metabolism Enzymes, Immobilized Epidermal Growth Factor / chemistry Epidermal Growth Factor / metabolism Female Heparin / metabolism Heparin-binding EGF-like Growth Factor Humans Intercellular Signaling Peptides and Proteins / chemistry Intercellular Signaling Peptides and Proteins / metabolism* Metalloproteases Mice Mice, Inbred BALB C Mice, Nude Microscopy, Confocal Neoplasm Invasiveness Protein Precursors / chemistry Protein Precursors / metabolism Protein Structure, Tertiary Transplants
IF 2.23
Times Cited 3
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
DNA material CSII-CMV-MCS-IRES2-Bsd (RDB04385)