title: [Cell model for the study of receptor and regulatory functions of human proHB-EGF].
abstract: Developing of new models and approaches, particularly with fluorescent techniques, for investigation of intracellular transport of proHB-EGF and its ligand-receptor complexes is strongly required. In order to create a model for studying proHB-EGF functions the genetic construction pEGFP-N1-proHB-EGF, encoding proHB-EGF-EGFP which is fluorescent-labeled form of proHB-EGF with enhanced green fluorescent protein EGFP in the cytoplasmic terminus of the molecule, was obtained. Eukaryotic cells expressing fusion protein proHB-EGF-EGFP on the cell surface were obtained by transfection with pEGFP-N1-proHB-EGF. Expressed in the Vero cells proHB-EGF-EGFP could bind fluorescent derivative of nontoxic receptor-binding subunit B of diphtheria toxin mCherry-SubB. After stimulation oftransfected cells with TPA (12-O-Tet-radecanoylphorbol-13-acetate), proHB-EGF-EGFP formed a fluorescentl-labeled C-terminal fragment of the molecule - CTF-EGFP. Thus, the obtained genetic construction pEGFP-N1-proHB-EGF could be helpful in visualization of molecules proHB-EGF and CTF in cells, may open new possibilities for the studying of their functions, such as receptor function of proHB-EGF for diphtheria toxin, intracellular translocation of CTF and provide possibilities for natural proHB-EGF ligands search.
authors.forename: N V
authors.initials: NV
authors.lastname: Korotkevych
authors.forename: A Iu
authors.initials: AIu
authors.lastname: Labyntsev
authors.forename: K Iu
authors.initials: KIu
authors.lastname: Manoĭlov
authors.forename: O I
authors.initials: OI
authors.lastname: Krynina
authors.forename: K V
authors.initials: KV
authors.lastname: Diachenko
authors.forename: D V
authors.initials: DV
authors.lastname: Kolybo
authors.forename: S V
authors.initials: SV
authors.lastname: Komisarenko
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D004167:Diphtheria Toxin / Q000235:genetics / Q000378:metabolism; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics; D066257:Heparin-binding EGF-like Growth Factor / Q000235:genetics* / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D008954:Models, Biological; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D014162:Transfection; D014709:Vero Cells; D000097573:Red Fluorescent Protein
chemical_list: D004167:Diphtheria Toxin; D066246:ErbB Receptors; D049452:Green Fluorescent Proteins; D066257:Heparin-binding EGF-like Growth Factor; D008164:Luminescent Proteins; D010446:Peptide Fragments; D000097573:Red Fluorescent Protein; C039171:diphtheria toxin fragment B; C485184:enhanced green fluorescent protein