Secreted glyceraldehye-3-phosphate dehydrogenase is a multifunctional autocrine transferrin receptor for cellular iron acquisition

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

doi:10.1016/j.bbagen.2013.03.019

Cited by 42 publications

(64 citation statements)

References 56 publications

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“…We selected GAPDH for further analysis because: (i) GAPDH has recently been identified as a transferrin receptor in macrophages and numerous other cell types [26][27][28] , (ii) the highly conserved nature of the protein and (iii) its role as a virulence factor [29][30][31][32] .…”

Section: Resultsmentioning

confidence: 99%

“…GAPDH has also been reported as a transferrin-binding protein in S. aureus and S. epidermidis 21 . In eukaryotes, numerous functions have been attributed to GAPDH including its role as a receptor for transferrin in different cell types and especially in macrophages [26][27][28]39 . Since this molecule is highly conserved across species (B50% identity with human GAPDH), we initially selected GAPDH to further explore the role of this transferrinbinding protein in M.tb.…”

Section: Discussionmentioning

confidence: 99%

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Mycobacterium tuberculosis acquires iron by cell-surface sequestration and internalization of human holo-transferrin

et al. 2014

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Mycobacterium tuberculosis (M.tb), which requires iron for survival, acquires this element by synthesizing iron-binding molecules known as siderophores and by recruiting a host iron-transport protein, transferrin, to the phagosome. The siderophores extract iron from transferrin and transport it into the bacterium. Here we describe an additional mechanism for iron acquisition, consisting of an M.tb protein that drives transport of human holo-transferrin into M.tb cells. The pathogenic strain M.tb H37Rv expresses several proteins that can bind human holo-transferrin. One of these proteins is the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Rv1436), which is present on the surface of M.tb and its relative Mycobacterium smegmatis. Overexpression of GAPDH results in increased transferrin binding to M.tb cells and iron uptake. Human transferrin is internalized across the mycobacterial cell wall in a GAPDH-dependent manner within infected macrophages.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Mycobacterium tuberculosis acquires iron by cell-surface sequestration and internalization of human holo-transferrin

et al. 2014

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“…CHO and THP1 cell lines were obtained from the National Centre for Cell Sciences, Pune, India. THP1 cells were activated through incubation with 12.5 ng/ml of phorbol 12-myristate 13-acetate (PMA) for 24 h. A stable THP1 cell line in which knockdown of total cellular GAPDH (by using GAPDH shRNA lentiviral particles) has been established previously (Sheokand et al, 2013) was used in the study. CHO-TRVb cells were silenced for GAPDH expression (by using GAPDH siRNA), as described previously (Kumar et al, 2012).…”

Section: Cell Lines Primary Cells and Materialsmentioning

confidence: 99%

“…The specific activity was typically ∼200 cpm/ng. Conjugates of apotransferrin with Alexa-Fluor-647, FITC and biotin were prepared exactly as described previously (Sheokand et al, 2013(Sheokand et al, , 2014.…”

Section: Radiolabeling Of Apotransferrinmentioning

confidence: 99%

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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“…It has also been reported that GAPDH is expressed on the surfaces of macrophages and functions as a transferrin or lactoferrin receptor in these cells [5,6] . Notably, macrophages have been demonstrated to secrete high levels of GAPDH [7] , indicating that secreted GAPDH could act as a regulatory factor during macrophage-mediated inflammation. However, the mechanisms underlying the secretion of GAPDH from macrophages have not been fully elucidated.…”

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ATP-triggered unconventional secretion of GAPDH from macrophages: Its putative role as a modulator of inflammation

et al. 2016

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Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key glycolytic enzyme that is predominantly localized in the cytoplasm. However, emerging evidence indicates that GAPDH is secreted from mammalian cells and performs some of its biological functions in the extracellular space. In particular, it has been reported that high levels of GAPDH are secreted from macrophages that play important roles in the innate immune system. Despite these findings, since GAPDH is a leaderless protein, the mechanisms by which it reaches the extracellular environment remain unclear. In this regard, we have recently reported that extracellular ATP is able to trigger the secretion of GAPDH from mouse microglial cells, the resident macrophages in the brain. In addition, the activation of microglial cells by lipopolysaccharide (LPS) clearly facilitated ATP-induced GAPDH secretion. Importantly, exosome secretion mediated by the P2X7 receptor (P2X7R), an ATP-gated cation channel, was shown to play a critical role in the induction of unconventional GAPDH secretion from LPS-primed microglial cells. We also found that secreted GAPDH affects the LPS-induced phosphorylation of p38 mitogen-activated protein kinase in microglial cells. Furthermore, our preliminary data demonstrated that the ATP-induced secretion of GAPDH occurs in macrophage cell lines derived from other mouse peripheral tissues, such as the liver. Together, our findings suggest that GAPDH is generated extracellularly through the stimulation of activated macrophages by ATP, and the secreted GAPDH acts as a mediator of macrophage-related inflammation in an autocrine/paracrine fashion.

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Secreted glyceraldehye-3-phosphate dehydrogenase is a multifunctional autocrine transferrin receptor for cellular iron acquisition

Cited by 42 publications

References 56 publications

Mycobacterium tuberculosis acquires iron by cell-surface sequestration and internalization of human holo-transferrin

Mycobacterium tuberculosis acquires iron by cell-surface sequestration and internalization of human holo-transferrin

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

ATP-triggered unconventional secretion of GAPDH from macrophages: Its putative role as a modulator of inflammation

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