3D Structure and Interaction of p24β and p24δ Golgi Dynamics Domains: Implication for p24 Complex Formation and Cargo Transport. Issue 20 (9th October 2016)

• Record Type: Journal Article
• Authors: Nagae, Masamichi; Hirata, Tetsuya; Morita-Matsumoto, Kana; Theiler, Romina; Fujita, Morihisa; Kinoshita, Taroh; Yamaguchi, Yoshiki
• Is Part Of: Journal of molecular biology. Volume 428:Issue 20(2016:Oct. 15)
• Page Start: 4087
• Page End: 4099
• Publication Date: 2016-10-09
• Subjects: TMED transmembrane Emp24 protein transport domain -- COP coat protein -- ER endoplasmic reticulum -- GOLD Golgi dynamics -- GPI-AP glycosylphosphatidylinositol-anchored protein -- myc-p24δ1 myc-tagged p24δ1 -- KO knockout -- VFG-GPI VSVGts-FLAG-mEGFP-GPI -- Dox doxycycline -- SAD single anomalous diffraction -- SeMet selenomethionine -- HSQC heteronuclear single quantum coherence -- Br Bromide; cargo receptor -- GOLD domain -- GPI-anchored protein -- p24 family -- protein transport; Molecular biology -- Periodicals; Biology -- Periodicals; Biochemistry -- Periodicals; Bacteriology -- Periodicals; Molecular Biology -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biologie -- Périodiques; Biochimie -- Périodiques; Moleculaire biologie; Biochemistry; Biology; Molecular biology; Periodicals; 572.805
• DOI: 10.1016/j.jmb.2016.08.023
• Languages: English
• ISSNs: 0022-2836

Abstract

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 ( K d = ~ 10 − 4 M). 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. Graphical Abstract: Highlights: p24 family proteins are involved in intracellular protein transport. p24δ1 GOLD domain is essential for efficient transport of glycosylphosphatidylinositol-anchored proteins. p24β1 and p24δ1 GOLD domains have a β-sandwich fold with an intrasheet disulfide bridge. p24β1 and p24δ1 GOLD domains interact weakly. Interaction of GOLD domains may contribute to p24 hetero-oligomeric complex formation.