Distinct EH domains of the endocytic TPLATE complex confer lipid and protein binding

Varování

Publikace nespadá pod Fakultu sportovních studií, ale pod Středoevropský technologický institut. Oficiální stránka publikace je na webu muni.cz.

Autoři	YPERMAN K. PAPAGEORGIOU Anna MERCERON R. DE MUNCK S. BLOCH Y. EECKHOUT D. JIANG Q.H. TACK P. GRIGORYAN R. EVANGELIDIS Thomas VAN LEENE J. VINCZE L. VANDENABEELE P. VANHAECKE F. POTOCKY M. DE JAEGER G. SAVVIDES S.N. TRIPSIANES Konstantinos PLESKOT R. VAN DAMME D.
Rok publikování	2021
Druh	Článek v odborném periodiku
Časopis / Zdroj	Nature Communications
Fakulta / Pracoviště MU	Středoevropský technologický institut
Citace
www	https://www.ncbi.nlm.nih.gov/pubmed/34031427
Doi	http://dx.doi.org/10.1038/s41467-021-23314-6
Klíčová slova	Adaptor Proteins; Signal Transducing; Calcium-Binding Proteins; Cell Membrane; X-Ray; NMR; Molecular Dynamics Simulation
Popis	Clathrin-mediated endocytosis (CME) is the gatekeeper of the plasma membrane. In contrast to animals and yeasts, CME in plants depends on the TPLATE complex (TPC), an evolutionary ancient adaptor complex. However, the mechanistic contribution of the individual TPC subunits to plant CME remains elusive. In this study, we used a multidisciplinary approach to elucidate the structural and functional roles of the evolutionary conserved N-terminal Eps15 homology (EH) domains of the TPC subunit AtEH1/Pan1. By integrating high-resolution structural information obtained by X-ray crystallography and NMR spectroscopy with all-atom molecular dynamics simulations, we provide structural insight into the function of both EH domains. Both domains bind phosphatidic acid with a different strength, and only the second domain binds phosphatidylinositol 4,5-bisphosphate. Unbiased peptidome profiling by mass-spectrometry revealed that the first EH domain preferentially interacts with the double N-terminal NPF motif of a previously unidentified TPC interactor, the integral membrane protein Secretory Carrier Membrane Protein 5 (SCAMP5). Furthermore, we show that AtEH/Pan1 proteins control the internalization of SCAMP5 via this double NPF peptide interaction motif. Collectively, our structural and functional studies reveal distinct but complementary roles of the EH domains of AtEH/Pan1 in plant CME and connect the internalization of SCAMP5 to the TPLATE complex. AtEH/Pan1 proteins contain two N-terminal Eps15 homology (EH) domains and are subunits of the endocytic TPLATE complex present in plants. Here, the authors combine X-ray crystallography, NMR and MD simulations with biochemical and in planta analysis to characterize the two AtEH1/Pan1 EH domains and reveal their structural differences and complementary functional roles.
Související projekty:	CEITEC 2020 Pushing the limits in automated NMR structure determination using a single 4D NOESY spectrum and machine learning methods