RT Journal Article
SR Electronic
T1 Cross-linking of the Endolysosomal System Reveals Flotillin Structures and Putative Cargo
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.01.12.475930
DO 10.1101/2022.01.12.475930
A1 Jasjot Singh
A1 Hadeer Elhabashy
A1 Pathma Muthukottiappan
A1 Markus Stepath
A1 Martin Eisenacher
A1 Oliver Kohlbacher
A1 Volkmar Gieselmann
A1 Dominic Winter
YR 2022
UL http://biorxiv.org/content/early/2022/01/12/2022.01.12.475930.abstract
AB Lysosomes are well-established as the main cellular organelles for the degradation of macromolecules and emerging as regulatory centers of metabolism. They are of crucial importance for cellular homeostasis, which is exemplified by a plethora of disorders related to alterations in lysosomal function. In this context, protein complexes play a decisive role, regulating not only metabolic lysosomal processes, but also lysosome biogenesis, transport, and interaction with other organelles. Using cross-linking mass spectrometry, we analyzed lysosomes and early endosomes. Based on the identification of 5,376 cross-links, we investigated protein-protein interactions and structures of lysosome- and endosome-related proteins. In particular, we present evidence for a tetrameric assembly of the lysosomal hydrolase PPT1 and heterodimeric/- multimeric structures of FLOT1/FLOT2 at lysosomes and early endosomes. For FLOT1-/FLOT2- positive early endosomes, we identified >300 proteins presenting putative cargo, and confirm the latrophilin family of adhesion G protein-coupled receptors as substrates for flotillin-dependent endocytosis.Competing Interest StatementThe authors have declared no competing interest.