Structural and functional insights into the interaction and targeting hub TMD0 of the polypeptide transporter TAPL

Abstract The ATP-binding cassette transporter TAPL translocates polypeptides from the cytosol into the lysosomal lumen. TAPL can be divided into two functional units: coreTAPL, active in ATP-dependent peptide translocation, and the N-terminal membrane spanning domain, TMD0, responsible for cellular...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Christoph Bock, Frank Löhr, Franz Tumulka, Katrin Reichel, Julia Würz, Gerhard Hummer, Lars Schäfer, Robert Tampé, Benesh Joseph, Frank Bernhard, Volker Dötsch, Rupert Abele
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Paramagnetic Relaxation Enhancement (PRE) Nuclear Overhauser Effects (NOEs) Helical Transmembrane (TMHs) Pulsed Electron-electron Double Resonance Nanodiscs Medicine R Science Q
Acceso en línea:	https://doaj.org/article/848161c3458d4b778156d6e149516c9c
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:848161c3458d4b778156d6e149516c9c
record_format	dspace
spelling	oai:doaj.org-article:848161c3458d4b778156d6e149516c9c2021-12-02T15:08:51ZStructural and functional insights into the interaction and targeting hub TMD0 of the polypeptide transporter TAPL10.1038/s41598-018-33841-w2045-2322https://doaj.org/article/848161c3458d4b778156d6e149516c9c2018-10-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-33841-whttps://doaj.org/toc/2045-2322Abstract The ATP-binding cassette transporter TAPL translocates polypeptides from the cytosol into the lysosomal lumen. TAPL can be divided into two functional units: coreTAPL, active in ATP-dependent peptide translocation, and the N-terminal membrane spanning domain, TMD0, responsible for cellular localization and interaction with the lysosomal associated membrane proteins LAMP-1 and LAMP-2. Although the structure and function of ABC transporters were intensively analyzed in the past, the knowledge about accessory membrane embedded domains is limited. Therefore, we expressed the TMD0 of TAPL via a cell-free expression system and confirmed its correct folding by NMR and interaction studies. In cell as well as cell-free expressed TMD0 forms oligomers, which were assigned as dimers by PELDOR spectroscopy and static light scattering. By NMR spectroscopy of uniformly and selectively isotope labeled TMD0 we performed a complete backbone and partial side chain assignment. Accordingly, TMD0 has a four transmembrane helix topology with a short helical segment in a lysosomal loop. The topology of TMD0 was confirmed by paramagnetic relaxation enhancement with paramagnetic stearic acid as well as by nuclear Overhauser effects with c6-DHPC and cross-peaks with water.Christoph BockFrank LöhrFranz TumulkaKatrin ReichelJulia WürzGerhard HummerLars SchäferRobert TampéBenesh JosephFrank BernhardVolker DötschRupert AbeleNature PortfolioarticleParamagnetic Relaxation Enhancement (PRE)Nuclear Overhauser Effects (NOEs)Helical Transmembrane (TMHs)Pulsed Electron-electron Double ResonanceNanodiscsMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-10 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Paramagnetic Relaxation Enhancement (PRE) Nuclear Overhauser Effects (NOEs) Helical Transmembrane (TMHs) Pulsed Electron-electron Double Resonance Nanodiscs Medicine R Science Q
spellingShingle	Paramagnetic Relaxation Enhancement (PRE) Nuclear Overhauser Effects (NOEs) Helical Transmembrane (TMHs) Pulsed Electron-electron Double Resonance Nanodiscs Medicine R Science Q Christoph Bock Frank Löhr Franz Tumulka Katrin Reichel Julia Würz Gerhard Hummer Lars Schäfer Robert Tampé Benesh Joseph Frank Bernhard Volker Dötsch Rupert Abele Structural and functional insights into the interaction and targeting hub TMD0 of the polypeptide transporter TAPL
description	Abstract The ATP-binding cassette transporter TAPL translocates polypeptides from the cytosol into the lysosomal lumen. TAPL can be divided into two functional units: coreTAPL, active in ATP-dependent peptide translocation, and the N-terminal membrane spanning domain, TMD0, responsible for cellular localization and interaction with the lysosomal associated membrane proteins LAMP-1 and LAMP-2. Although the structure and function of ABC transporters were intensively analyzed in the past, the knowledge about accessory membrane embedded domains is limited. Therefore, we expressed the TMD0 of TAPL via a cell-free expression system and confirmed its correct folding by NMR and interaction studies. In cell as well as cell-free expressed TMD0 forms oligomers, which were assigned as dimers by PELDOR spectroscopy and static light scattering. By NMR spectroscopy of uniformly and selectively isotope labeled TMD0 we performed a complete backbone and partial side chain assignment. Accordingly, TMD0 has a four transmembrane helix topology with a short helical segment in a lysosomal loop. The topology of TMD0 was confirmed by paramagnetic relaxation enhancement with paramagnetic stearic acid as well as by nuclear Overhauser effects with c6-DHPC and cross-peaks with water.
format	article
author	Christoph Bock Frank Löhr Franz Tumulka Katrin Reichel Julia Würz Gerhard Hummer Lars Schäfer Robert Tampé Benesh Joseph Frank Bernhard Volker Dötsch Rupert Abele
author_facet	Christoph Bock Frank Löhr Franz Tumulka Katrin Reichel Julia Würz Gerhard Hummer Lars Schäfer Robert Tampé Benesh Joseph Frank Bernhard Volker Dötsch Rupert Abele
author_sort	Christoph Bock
title	Structural and functional insights into the interaction and targeting hub TMD0 of the polypeptide transporter TAPL
title_short	Structural and functional insights into the interaction and targeting hub TMD0 of the polypeptide transporter TAPL
title_full	Structural and functional insights into the interaction and targeting hub TMD0 of the polypeptide transporter TAPL
title_fullStr	Structural and functional insights into the interaction and targeting hub TMD0 of the polypeptide transporter TAPL
title_full_unstemmed	Structural and functional insights into the interaction and targeting hub TMD0 of the polypeptide transporter TAPL
title_sort	structural and functional insights into the interaction and targeting hub tmd0 of the polypeptide transporter tapl
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/848161c3458d4b778156d6e149516c9c
work_keys_str_mv	AT christophbock structuralandfunctionalinsightsintotheinteractionandtargetinghubtmd0ofthepolypeptidetransportertapl AT franklohr structuralandfunctionalinsightsintotheinteractionandtargetinghubtmd0ofthepolypeptidetransportertapl AT franztumulka structuralandfunctionalinsightsintotheinteractionandtargetinghubtmd0ofthepolypeptidetransportertapl AT katrinreichel structuralandfunctionalinsightsintotheinteractionandtargetinghubtmd0ofthepolypeptidetransportertapl AT juliawurz structuralandfunctionalinsightsintotheinteractionandtargetinghubtmd0ofthepolypeptidetransportertapl AT gerhardhummer structuralandfunctionalinsightsintotheinteractionandtargetinghubtmd0ofthepolypeptidetransportertapl AT larsschafer structuralandfunctionalinsightsintotheinteractionandtargetinghubtmd0ofthepolypeptidetransportertapl AT roberttampe structuralandfunctionalinsightsintotheinteractionandtargetinghubtmd0ofthepolypeptidetransportertapl AT beneshjoseph structuralandfunctionalinsightsintotheinteractionandtargetinghubtmd0ofthepolypeptidetransportertapl AT frankbernhard structuralandfunctionalinsightsintotheinteractionandtargetinghubtmd0ofthepolypeptidetransportertapl AT volkerdotsch structuralandfunctionalinsightsintotheinteractionandtargetinghubtmd0ofthepolypeptidetransportertapl AT rupertabele structuralandfunctionalinsightsintotheinteractionandtargetinghubtmd0ofthepolypeptidetransportertapl
_version_	1718388009504604160

Structural and functional insights into the interaction and targeting hub TMD0 of the polypeptide transporter TAPL

Ejemplares similares