title: Production, Isolation, and Structural Analysis of Ligands and Receptors of the TGF-β Superfamily.
abstract: The ability to understand the molecular mechanisms by which secreted signaling proteins of the TGF-β superfamily assemble their cell surface receptors into complexes to initiate downstream signaling is dependent upon the ability to determine atomic-resolution structures of the signaling proteins, the ectodomains of the receptors, and the complexes that they form. The structures determined to date have revealed major differences in the overall architecture of the signaling complexes formed by the TGF-βs and BMPs, which has provided insights as to how they have evolved to fulfill their distinct functions. Such studies, have however, only been applied to a few members of the TGF-β superfamily, which is largely due to the difficulty of obtaining milligram-scale quantities of highly homogenous preparations of the disulfide-rich signaling proteins and receptor ectodomains of the superfamily. Here we describe methods used to produce signaling proteins and receptor ectodomains of the TGF-β superfamily using bacterial and mammalian expression systems and procedures to purify them to homogeneity.
authors.affiliation: Protein Chemistry, Novo Nordisk Research Center China, 20 Life Science Park Rd, Bldg 2, Beijing, 102206, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Huang
authors.affiliation: Protein Chemistry, Novo Nordisk Research Center China, 20 Life Science Park Rd, Bldg 2, Beijing, 102206, China. ahinck@pitt.edu.
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Hinck
mesh_terms: D000818:Animals; D002460:Cell Line; D018360:Crystallography, X-Ray; D015870:Gene Expression; D006801:Humans; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D018125:Receptors, Transforming Growth Factor beta / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins; D015398:Signal Transduction; D016212:Transforming Growth Factor beta / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D008024:Ligands; D018125:Receptors, Transforming Growth Factor beta; D011993:Recombinant Fusion Proteins; D016212:Transforming Growth Factor beta
keywords: TGF-β; ligand and receptor; protein expression; protein purification; structural analysis