title: Erythropoietin derived by chemical synthesis.
abstract: Erythropoietin is a signaling glycoprotein that controls the fundamental process of erythropoiesis, orchestrating the production and maintenance of red blood cells. As administrated clinically, erythropoietin has a polypeptide backbone with complex dishomogeneity in its carbohydrate domains. Here we describe the total synthesis of homogeneous erythropoietin with consensus carbohydrate domains incorporated at all of the native glycosylation sites. The oligosaccharide sectors were built by total synthesis and attached stereospecifically to peptidyl fragments of the wild-type primary sequence, themselves obtained by solid-phase peptide synthesis. The glycopeptidyl constructs were joined by chemical ligation, followed by metal-free dethiylation, and subsequently folded. This homogeneous erythropoietin glycosylated at the three wild-type aspartates with N-linked high-mannose sialic acid-containing oligosaccharides and O-linked glycophorin exhibits Procrit-level in vivo activity in mice.
authors: Laboratory for Bioorganic Chemistry, Sloan-Kettering Institute for Cancer Research, 1275 York Avenue, New York, NY 10065, USA.
authors: Ping
authors: P
authors: Wang
authors: Laboratory for Bioorganic Chemistry, Sloan-Kettering Institute for Cancer Research, 1275 York Avenue, New York, NY 10065, USA.
authors: Suwei
authors: S
authors: Dong
authors: Cell Biology Program, Sloan-Kettering Institute for Cancer Research, 1275 York Avenue, New York, NY 10065, USA.
authors: Jae-Hung
authors: JH
authors: Shieh
authors: Cell Biology Program, Sloan-Kettering Institute for Cancer Research, 1275 York Avenue, New York, NY 10065, USA.
authors: Elizabeth
authors: E
authors: Peguero
authors: Department of Pharmacology and Chemistry, Sloan-Kettering Institute for Cancer Research, 1275 York Avenue, New York, NY 10065, USA.
authors: Ronald
authors: R
authors: Hendrickson
authors: Cell Biology Program, Sloan-Kettering Institute for Cancer Research, 1275 York Avenue, New York, NY 10065, USA.
authors: Malcolm A S
authors: MAS
authors: Moore
authors: Laboratory for Bioorganic Chemistry, Sloan-Kettering Institute for Cancer Research, 1275 York Avenue, New York, NY 10065, USA.|Department of Chemistry, Columbia University, Havemeyer Hall, 3000 Broadway, New York, NY 10027, USA.
authors: Samuel J
authors: SJ
authors: Danishefsky
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001224:Aspartic Acid / Q000737:chemistry; D002478:Cells, Cultured; D016384:Consensus Sequence; D004305:Dose-Response Relationship, Drug; D004906:Erythrocyte Count; D004921:Erythropoietin / Q000008:administration & dosage* / Q000138:chemical synthesis* / Q000737:chemistry; D006021:Glycophorins / Q000737:chemistry; D006031:Glycosylation; D007279:Injections, Subcutaneous; D008358:Mannose / Q000737:chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry; D012156:Reticulocytes / Q000187:drug effects
chemical_list: D006021:Glycophorins; D009844:Oligosaccharides; D004921:Erythropoietin; D001224:Aspartic Acid; D019158:N-Acetylneuraminic Acid; D008358:Mannose