title: The diphtheria toxin transmembrane domain as a pH sensitive membrane anchor for human interleukin-2 and murine interleukin-3.
abstract: We have constructed two fusion proteins T-hIL-2 and T-mIL-3 in which human interleukin-2 (hIL-2) or murine interleukin-3 (mIL-3) are fused to the C-terminus of the diphtheria toxin transmembrane domain (T domain). Two additional fusion proteins, T-(Gly4-Ser)2-hIL-2 and T-(Gly4-Ser)2-mIL-3, were derived by introduction of the (Gly4-Ser)2 spacer between the T domain and cytokine components. Recognition of the hIL-2 receptor or the mIL-3 receptor by the corresponding recombinant proteins was demonstrated by their capacity to stimulate cytokine-dependent cell lines. All proteins retained the capacity of the T domain to insert into phospholipid membranes at acidic pH. Finally, anchoring of both cytokines to the membrane of lipid vesicles or living cells was assessed by specific antibody recognition. Our results show that the T domain fused to the N-terminus of a given protein can function as a pH sensitive membrane anchor for that protein.
authors.affiliation: Département d'Ingénierie et d'Etudes des Protéines, DSV, CEA, CE, Gif-sur-Yvette, France.
authors.forename: D
authors.initials: D
authors.lastname: Liger
authors.forename: P
authors.initials: P
authors.lastname: Nizard
authors.forename: C
authors.initials: C
authors.lastname: Gaillard
authors.forename: J C
authors.initials: JC
authors.lastname: vanderSpek
authors.forename: J R
authors.initials: JR
authors.lastname: Murphy
authors.forename: B
authors.initials: B
authors.lastname: Pitard
authors.forename: D
authors.initials: D
authors.lastname: Gillet
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002462:Cell Membrane / Q000378:metabolism; D017931:DNA Primers; D004167:Diphtheria Toxin / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007376:Interleukin-2 / Q000378:metabolism*; D007377:Interleukin-3 / Q000378:metabolism*; D008081:Liposomes; D051379:Mice; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D017931:DNA Primers; D004167:Diphtheria Toxin; D007376:Interleukin-2; D007377:Interleukin-3; D008081:Liposomes; D011993:Recombinant Fusion Proteins