Connecting the Dots in the Neuroglobin-Protein Interaction Network of an Unstressed and Ferroptotic Cell Death Neuroblastoma Model

Abstract

Neuroglobin is a heme protein of which increased levels provide neuroprotection against amyloid proteinopathy and hemorrhagic damage. These cellular stressors involve the promotion of ferroptosis—an iron-dependent, lipid peroxide-accreting form of cell death. Hence, we questioned whether neuroglobin could oppose ferroptosis initiation. We detected human neuroglobin (hNgb)-EGFP-expressing SH-SY5Y cells to be significantly more resistant to ferroptosis induction, identifying 0.68-fold less cell death. To elucidate the underlying pathways, this study investigated hNgb-protein interactions with a Co-IP-MS/MS approach both under a physiological and a ferroptotic condition. hNgb binds to proteins of the cellular iron metabolism (e.g., RPL15 and PCBP3) in an unstressed condition and shows an elevated binding ratio towards cell death-linked proteins, such as HNRNPA3, FAM120A, and ABRAXAS2, under ferroptotic stress. Our data also reveal a constitutive interaction between hNgb and the longevity-associated heterodimer XRCC5/XRCC6. Disentangling the involvement of hNgb and its binding partners in cellular processes, using Ingenuity Pathway Analysis, resulted in the integration of hNgb in the ubiquitination pathway, mTOR signaling, 14-3-3-mediated signaling, and the glycolysis cascade. We also detected a previously unknown strong link with motor neuropathies. Hence, this study contributes to the elucidation of neuroglobin’s involvement in cellular mechanisms that provide neuroprotection and the upkeep of homeostasis.

The role of ferroptosis in breast cancer patients: a comprehensive analysis

“…G6PD is associated with mitochondrial dysfunction 26 and human neuroglobin. It binds proteins of the cellular iron metabolism under ferroptosis stress 27 . Prominin2 ( PROM2 ) can suppress ferroptosis sensitivity 28 .…”

CD56 Homodimerization and Participation in Anti-Tumor Immune Effector Cell Functioning: A Role for Interleukin-15

“…The acute myeloid leukemia cell line NB4 was purchased from the Deutsche Sammlung von Mikroorganismen und Zellkulturen (Braunschweig, Germany). The SH-SY5Y and SH-SY5Y-eGFP [61] neuroblastoma cell lines were kindly provided to us by the laboratory of Prof. Dewilde (University of Antwerp, Belgium) and the multiple myeloma cell line U266 was a gift from Prof. Germeraad (Maastricht University, The Netherlands). Peripheral blood mononuclear cells were isolated from buffy coats of healthy donors, provided by the Red Cross donor center (Red Cross-Flanders, Mechelen, Belgium).…”

hnRNP A/B Proteins: An Encyclopedic Assessment of Their Roles in Homeostasis and Disease

“…Gilpin et al [ 293 ] found that interaction with Ubiquilin 2 stabilized A1 to maintain protein abundance, and presume that it would have a similar effect on A3, providing a connection to the other pathological A3 observations above. Tangentially, van Acker et al [ 294 ] recently identified an interaction between A3 and neuroglobin, a neuroprotective protein that prevents ferroptosis in response to proteinopathy. During ferroptotic stress, neuroglobin interacts with A3, but the outcome of this interaction is unknown [ 294 ].…”

“…Tangentially, van Acker et al [ 294 ] recently identified an interaction between A3 and neuroglobin, a neuroprotective protein that prevents ferroptosis in response to proteinopathy. During ferroptotic stress, neuroglobin interacts with A3, but the outcome of this interaction is unknown [ 294 ]. Altogether, these findings suggest a greater role for A3 in ALS pathogenesis than just HR and DPR regulation.…”