title: 3D Structure and Interaction of p24β and p24δ Golgi Dynamics Domains: Implication for p24 Complex Formation and Cargo Transport.
abstract: The p24 family consists of four subfamilies (p24α, p24β, p24γ, and p24δ), and the proteins are thought to form hetero-oligomeric complexes for efficient transport of cargo proteins from the endoplasmic reticulum to the Golgi apparatus. The proteins possess a conserved luminal Golgi dynamics (GOLD) domain, whose functions are largely unknown. Here, we present structural and biochemical studies of p24β1 and p24δ1 GOLD domains. Use of GOLD domain-deleted mutants revealed that the GOLD domain of p24δ1 is required for proper p24 hetero-oligomeric complex formation and efficient transport of GPI-anchored proteins. The p24β1 and p24δ1 GOLD domains share a common β-sandwich fold with a characteristic intrasheet disulfide bond. The GOLD domain of p24δ1 crystallized as dimers, allowing the analysis of a homophilic interaction site. Surface plasmon resonance and solution NMR analyses revealed that p24β1 and p24δ1 GOLD domains interact weakly (Kd= ~10-4M). Bi-protein titration provided interaction site maps. We propose that the heterophilic interaction of p24 GOLD domains contributes to the formation of the p24 hetero-oligomeric complex and to efficient cargo transport.
authors: Structural Glycobiology Team, Systems Glycobiology Research Group, RIKEN-Max Planck Joint Research Center, RIKEN Global Research Cluster, 2-1 Hirosawa, Wako City, Saitama 351-0198, Japan.
authors: Masamichi
authors: M
authors: Nagae
authors: Research Institute for Microbial Diseases and WPI Immunology Frontier Research Center, Osaka University, Suita, Osaka 565-0871, Japan.
authors: Tetsuya
authors: T
authors: Hirata
authors: Structural Glycobiology Team, Systems Glycobiology Research Group, RIKEN-Max Planck Joint Research Center, RIKEN Global Research Cluster, 2-1 Hirosawa, Wako City, Saitama 351-0198, Japan.
authors: Kana
authors: K
authors: Morita-Matsumoto
authors: Research Institute for Microbial Diseases and WPI Immunology Frontier Research Center, Osaka University, Suita, Osaka 565-0871, Japan.
authors: Romina
authors: R
authors: Theiler
authors: Research Institute for Microbial Diseases and WPI Immunology Frontier Research Center, Osaka University, Suita, Osaka 565-0871, Japan; Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, China.
authors: Morihisa
authors: M
authors: Fujita
authors: Research Institute for Microbial Diseases and WPI Immunology Frontier Research Center, Osaka University, Suita, Osaka 565-0871, Japan.
authors: Taroh
authors: T
authors: Kinoshita
authors: Structural Glycobiology Team, Systems Glycobiology Research Group, RIKEN-Max Planck Joint Research Center, RIKEN Global Research Cluster, 2-1 Hirosawa, Wako City, Saitama 351-0198, Japan. Electronic address: yyoshiki@riken.jp.
authors: Yoshiki
authors: Y
authors: Yamaguchi
mesh_terms: D002460:Cell Line; D018360:Crystallography, X-Ray; D006056:Golgi Apparatus / Q000378:metabolism*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D029543:Nucleocytoplasmic Transport Proteins; D011485:Protein Binding; D011487:Protein Conformation; D025941:Protein Interaction Mapping; D055503:Protein Multimerization; D021381:Protein Transport; D020349:Surface Plasmon Resonance; D033921:Vesicular Transport Proteins
chemical_list: D008565:Membrane Proteins; D029543:Nucleocytoplasmic Transport Proteins; C484395:TMED10 protein, human; C524397:TMED2 protein, human; D033921:Vesicular Transport Proteins
keywords: GOLD domain; GPI-anchored protein; cargo receptor; p24 family; protein transport