Moonlighting cell surface GAPDH recruits Apo Transferrin to effect iron egress from mammalian cells

Sheokand, Navdeep; Malhotra, Himanshu; Kumar, Santosh; Tillu, Vikas A.; Chauhan, Anoop Singh; Raje, Chaaya Iyengar; Raje, Manoj

doi:10.1242/jcs.154005

Cited by 35 publications

(54 citation statements)

References 56 publications

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“…1C). This matches well with our earlier reported value of 1.1 nM for apotransferrin binding of GAPDH on J774 cells, it is also similar to the K d of 5.3 nM reported by us for the in vitro GAPDH-apotransferrin interaction (Sheokand et al, 2014). Co-immunoprecipitation analysis confirmed the interaction of apotransferrin with surface GAPDH (Fig.…”

Section: Resultssupporting

confidence: 92%

“…1D). Our earlier work has also established the role of GAPDH in apotransferrin-facilitated Fe export from macrophages and hepatocytes (Sheokand et al, 2014), and here too we observed that knockdown of GAPDH (see Materials and Methods) significantly compromised the ability of Fe-loaded CHO-TRVb cells to export Fe in the presence of apotransferrin ( Fig. 1E; Fig.…”

Section: Resultssupporting

confidence: 80%

“…Controls were set up in parallel with normal medium. No significant change in viability was observed, as assessed with Trypan Blue, sulforhodamine and propidium iodide assays, as described previously (Sheokand et al, 2014).…”

Section: Cell Treatmentmentioning

confidence: 96%

“…Apotransferrin binding was assessed by using 10 µg apotransferrin conjugated to Alexa-Fluor-647. The incubation buffer for all apotransferrin-binding experiments included 100 µM desferrioxamine (DFO) to prevent incorporation of any Fe into apotransferrin (Kawabata et al, 2000;Sheokand et al, 2014). For intracellular staining, cells were fixed with 2% paraformaldehyde and treated with 0.1% saponin before incubation with antibodies.…”

Section: Flow Cytometry Analysismentioning

confidence: 99%

“…This isoform of GAPDH captures apotransferrin with high affinity and enhances cellular Fe export. The model proposed for this process involves Fe extrusion from cells through the classic cell surface ferroportin pump, followed by rapid chelation of the externalized Fe into apotransferrin (Sheokand et al, 2014). These findings still cannot explain as to how the vast majority of cells that lack ferroportin on their surface are able to extrude excess of the metal ions (D'Anna et al, 2009;Donovan et al, 2005), and the possibility that GAPDH utilizes an alternative method to remove Fe from cells in a surface-ferroportinindependent manner cannot be ruled out.…”

Section: Introductionmentioning

confidence: 99%

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Reverse overshot water-wheel retroendocytosis of Apo Transferrin extrudes cellular iron

Sheokand

Malhotra

Chauhan

et al. 2016

Journal of Cell Science

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Iron a vital micronutrient for all organisms must be managed judiciously as both, deficiency or excess can trigger severe pathology. While cellular iron import is well understood its export is thought to be limited to transmembrane extrusion via ferroportin the only known mammalian iron exporter. Utilizing primary cells and cell lines (including those with no discernible expression of ferroportin on their surface) we demonstrate that upon iron loading the multifunctional enzyme Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) that is recruited to the cell surface treadmills apo transferrin (apo Tf) in and out of the cell. Kinetic analysis utilizing; labeled ligand, GAPDH knock down cells, Fe55 labeled cells and pharmacological inhibitors of endocytosis confirmed GAPDH dependent apo Tf internalization as a prerequisite for cellular iron export. These studies define an unusual rapid recycling process of retroendocytosis for cellular iron extrusion, a process mirroring receptor mediated internalization that has never before been considered for maintenance of cellular cationic homeostasis. Modulation of this unusual pathway could provide insights for management of iron overload disorders.

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Section: Resultssupporting

confidence: 92%

Section: Resultssupporting

confidence: 80%

Section: Cell Treatmentmentioning

confidence: 96%

Section: Flow Cytometry Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Reverse overshot water-wheel retroendocytosis of Apo Transferrin extrudes cellular iron

Sheokand

Malhotra

Chauhan

et al. 2016

Journal of Cell Science

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Dimethyl fumarate, a two‐edged drug: Current status and future directions

Saidu

Kavian

Leroy

et al. 2019

Medicinal Research Reviews

120

100

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Dimethyl fumarate (DMF) is a fumaric acid ester registered for the treatment of relapsing‐remitting multiple sclerosis (RRMS). It induces protein succination leading to inactivation of cysteine‐rich proteins. It was first shown to possess cytoprotective and antioxidant effects in noncancer models, which appeared related to the induction of the nuclear factor erythroid 2 (NF‐E2)–related factor 2 (NRF2) pathway. DMF also displays antitumor activity in several cellular and mice models. Recently, we showed that the anticancer mechanism of DMF is dose‐dependent and is paradoxically related to the decrease in the nuclear translocation of NRF2. Some other studies performed indicate also the potential role of DMF in cancers, which are dependent on the NRF2 antioxidant and cellular detoxification program, such as KRAS‐mutated lung adenocarcinoma. It, however, seems that DMF has multiple biological effects as it has been shown to also inhibit the transcription factor nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐κB), thus blocking downstream targets that may be involved in the development and progression of inflammatory cascades leading to various disease processes, including tumors, lymphomas, diabetic retinopathy, arthritis, and psoriasis. Herein, we present the current status and future directions of the use of DMF in various diseases models with particular emphases on its targeting of specific intracellular signal transduction cascades in cancer; to shed some light on its possible mode of action.

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D-Glyceraldehyde-3-Phosphate Dehydrogenase Structure and Function

White

Garcin

2017

Subcellular Biochemistry

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Aside from its well-established role in glycolysis, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has been shown to possess many key functions in cells. These functions are regulated by protein oligomerization , biomolecular interactions, post-translational modifications , and variations in subcellular localization . Several GAPDH functions and regulatory mechanisms overlap with one another and converge around its role in intermediary metabolism. Several structural determinants of the protein dictate its function and regulation. GAPDH is ubiquitously expressed and is found in all domains of life. GAPDH has been implicated in many diseases, including those of pathogenic, cardiovascular, degenerative, diabetic, and tumorigenic origins. Understanding the mechanisms by which GAPDH can switch between its functions and how these functions are regulated can provide insights into ways the protein can be modulated for therapeutic outcomes.

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Moonlighting cell surface GAPDH recruits Apo Transferrin to effect iron egress from mammalian cells

Cited by 35 publications

References 56 publications

Reverse overshot water-wheel retroendocytosis of Apo Transferrin extrudes cellular iron

Reverse overshot water-wheel retroendocytosis of Apo Transferrin extrudes cellular iron

Dimethyl fumarate, a two‐edged drug: Current status and future directions

D-Glyceraldehyde-3-Phosphate Dehydrogenase Structure and Function

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