title: Engineering and use of (32)P-labeled human recombinant interleukin-11 for receptor binding studies.
abstract: Human interleukin-11 (hIL-11) is a pleiotropic cytokine that is involved in numerous biological activities such as hematopoiesis, osteoclastogenesis, neurogenesis and female fertility. IL-11 is obviously a key reagent to study the IL-11 receptors. However, conventional radio-iodination techniques lead to a loss of IL-11 bioactivity. Here, we report the construction and the production of a new recombinant human IL-11 (FP Delta IL-11). In this molecule, a specific phosphorylation site (RRASVA) has been introduced at the N-terminus of rhIL-11. It can be specifically phosphorylated by bovine heart protein kinase and accordingly, easily radiolabeled with (32)P. A high radiological specific activity (250,000 c.p.m x ng(-1) of protein) was obtained with the retention of full biological activity of the protein. The binding of (32)P-labeled FP Delta IL-11 to Ba/F3 cells stably transfected with plasmids encoding human IL-11 receptors alpha and beta chains (IL-11R alpha and gp130) was specific and saturable with a high affinity as determined from Scatchard plot analysis. Availability of this new ligand should prompt further studies on IL-11R structure, expression and regulation.
authors.affiliation: Institut Pasteur de Bruxelles, Belgium.
authors.forename: Xiao-Ming
authors.initials: XM
authors.lastname: Wang
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Wilkin
authors.forename: Olivier
authors.initials: O
authors.lastname: Boisteau
authors.forename: Dimitri
authors.initials: D
authors.lastname: Harmegnies
authors.forename: Chrystel
authors.initials: C
authors.lastname: Blanc
authors.forename: Paul
authors.initials: P
authors.lastname: Vandenbussche
authors.forename: Félix A
authors.initials: FA
authors.lastname: Montero-Julian
authors.forename: Yannick
authors.initials: Y
authors.lastname: Jacques
authors.forename: Jean
authors.initials: J
authors.lastname: Content
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D002478:Cells, Cultured; D006801:Humans; D017370:Interleukin-11 / Q000378:metabolism*; D008969:Molecular Sequence Data; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D015202:Protein Engineering; D018121:Receptors, Cytokine / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D017370:Interleukin-11; D010761:Phosphorus Radioisotopes; D018121:Receptors, Cytokine; D011994:Recombinant Proteins