Cathepsin D interacts with adenosine A2A receptors in mouse macrophages to modulate cell surface localization and inflammatory signaling.

Abstract

Adenosine A2A receptor (A2AR)-dependent signaling in macrophages plays a key role in the regulation of inflammation. However, the processes regulating A2AR targeting to the cell surface and degradation in macrophages are incompletely understood. For example, the C-terminal domain of the A2AR and proteins interacting with it are known to regulate receptor recycling, although it is unclear what role potential A2AR-interacting partners have in macrophages. Here, we aimed to identify A2AR-interacting partners in macrophages that may effect receptor trafficking and activity. To this end, we performed a yeast two-hybrid screen using the C-terminal tail of A2AR as the "bait" and a macrophage expression library as the "prey." We found that the lysosomal protease cathepsin D (CtsD) was a robust hit. The A2AR-CtsD interaction was validated in vitro and in cellular models, including RAW 264.7 and mouse peritoneal macrophage (IPMΦ) cells. We also demonstrated that the A2AR is a substrate of CtsD and that the blockade of CtsD activity increases the density and cell surface targeting of A2AR in macrophages. Conversely, we demonstrate that A2AR activation prompts the maturation and enzymatic activity of CtsD in macrophages. In summary, we conclude that CtsD is a novel A2AR-interacting partner and thus describe molecular and functional interplay that may be crucial for adenosine-mediated macrophage regulation in inflammatory processes.

• Authors: Adrienn Skopál, Tamás Kéki, Péter Á Tóth, Balázs Csóka, Balázs Koscsó, Zoltán H Németh, Luca Antonioli, Andreas Ivessa, Francisco Ciruela, László Virág, György Haskó, Endre Kókai
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 298 Issue 5 Pg. 101888 (05 2022) ISSN: 1083-351X [Electronic] United States
• PMID: 35367412 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.

Chemical References

• Carrier Proteins
• Receptor, Adenosine A2A
• Cathepsin D
• Ctsd protein, mouse
• Adenosine

Topics

• Adenosine (metabolism)
• Animals
• Carrier Proteins (metabolism)
• Cathepsin D (genetics, metabolism)
• Macrophages (metabolism)
• Mice
• Receptor, Adenosine A2A (genetics, metabolism)
• Signal Transduction