2.A.97 The Mitochondrial Inner Membrane K+/H+ and Ca2+/H+ Exchanger (LetM1) Family

Mitochondria are integral components of cellular calcium (Ca2+) signaling. Calcium stimulates mitochondrial adenosine 5''-triphosphate production, but can also initiate apoptosis. In turn, cytoplasmic Ca2+ concentrations are regulated by mitochondria. Leucine zipper EF hand-containing transmembrane protein 1 (LetM1) and uncoupling proteins 2 and 3 (UCP2/3) contribute to two distinct mitochondrial Ca2+ uptake pathways (Waldeck-Weiermair et al., 2011).  Lin and Stathopulos 2019 presented an overview of the current understanding of LETM1 structure, mechanism and function. LETM1 is required for mitochondrial homeostasis and cellular viability (Li et al. 2019).

LetM1 is a large human protein of 739 aas with 1 TMS. It is implicated in the Wolf-Hirschhorn Syndrome. Homologues are found in plants and fungi. These proteins in yeast and humans are in the inner mitochondrial membrane and are complementary. The human protein corrects the yeast defect. Deletion of the yeast protein yields defective, swollen mitochondria with increased cation (K+) contents and low Δψ. The primary defect may be in the mitochondrial K+/H+ exchange activity. These proteins therefore function both in K+ homeostasis and organelle volume control (Nowikovsky et al., 2004).

Jiang et al. (2009) searched for genes that regulate mitochondrial Ca2+ and H+ concentrations using a genome-wide Drosophila RNA interference (RNAi) screen. The mammalian homolog of one Drosophila gene identified in the screen, LetM1, was found to specifically mediate coupled Ca2+/H+ exchange. RNAi knockdown, overexpression, and liposome reconstitution of the purified LetM1 protein demonstrate that LetM1 is a mitochondrial Ca2+/H+ antiporter (Jiang et al., 2009). This system is discussed by Santo-Domingo and Demaurex (2010)

Leucine zipper-EF-hand containing transmembrane protein 1 (LetM1), one of the genes deleted in Wolf-Hirschhorn syndrome, encodes a putative mitochondrial Ca2+/H+ antiporter. Cellular Letm1 knockdown reduced Ca2+mito uptake, H+mito extrusion and impaired mitochondrial ATP generation capacity. Homozygous deletion of LetM1 in mice resulted in embryonic lethality before day 6.5 of embryogenesis and ~50% of the heterozygotes died before day 13.5 of embryogenesis (Jiang et al. 2013). The surviving heterozygous mice exhibited altered glucose metabolism, impaired control of brain ATP levels, and increased seizure activity. Thus, loss of LetM1 contributes to the pathology of Wolf-Hirschhorn syndrome in humans and may contribute to seizure phenotypes by reducing glucose oxidation.

Purified, and reconstituted human LetM1 exhibits apparent affinities of cations in the order Ca2+ = Mn2+ > Gd3+ = La3+ > Sr2+ >> Ba2+, Mg2+, K+, Na+. Kinetic analyses suggested a LetM1 turnover rate of 2 Ca2+/s and a Km of ~ 25 microM. LetM1 mediates electroneutral 1 Ca2+/2 H+ antiport. LetM1 is insensitive to ruthenium red, an inhibitor of the mitochondrial calcium uniporter, and CGP-37157, an inhibitor of the mitochondrial Na+/Ca2+exchanger. Functional properties of LetM1 are similar to those of the H+-dependent Ca2+ transport mechanism identified in intact mitochondria (Tsai et al. 2013).

This family belongs to the .



Alam, M.R., L.N. Groschner, W. Parichatikanond, L. Kuo, A.I. Bondarenko, R. Rost, M. Waldeck-Weiermair, R. Malli, and W.F. Graier. (2012). Mitochondrial Ca2+ uptake 1 (MICU1) and mitochondrial ca2+ uniporter (MCU) contribute to metabolism-secretion coupling in clonal pancreatic β-cells. J. Biol. Chem. 287: 34445-34454.

De Marchi, U., J. Santo-Domingo, C. Castelbou, I. Sekler, A. Wiederkehr, and N. Demaurex. (2014). NCLX protein, but not LETM1, mediates mitochondrial Ca2+ extrusion, thereby limiting Ca2+-induced NAD(P)H production and modulating matrix redox state. J. Biol. Chem. 289: 20377-20385.

Jiang, D., L. Zhao, and D.E. Clapham. (2009). Genome-wide RNAi screen identifies Letm1 as a mitochondrial Ca2+/H+ antiporter. Science 326: 144-147.

Jiang, D., L. Zhao, C.B. Clish, and D.E. Clapham. (2013). Letm1, the mitochondrial Ca2+/H+ antiporter, is essential for normal glucose metabolism and alters brain function in Wolf-Hirschhorn syndrome. Proc. Natl. Acad. Sci. USA 110: E2249-2254.

Li, Y., Q. Tran, R. Shrestha, L. Piao, S. Park, J. Park, and J. Park. (2019). LETM1 is required for mitochondrial homeostasis and cellular viability (Review). Mol Med Rep 19: 3367-3375.

Lin, Q.T. and P.B. Stathopulos. (2019). Molecular Mechanisms of Leucine Zipper EF-Hand Containing Transmembrane Protein-1 Function in Health and Disease. Int J Mol Sci 20:.

Nowikovsky, K., E.M. Froschauer, G. Zsurka, J. Samaj, S. Reipert, M. Kolisek, G. Wiesenberger, and R.J. Schweyen. (2004). The LETM1/YOL027 gene family encodes a factor of the mitochondrial K+ homeostasis with a potential role in the Wolf-Hirschhorn syndrome. J. Biol. Chem. 279: 30307-30315.

Santo-Domingo J. and Demaurex N. (2010). Calcium uptake mechanisms of mitochondria. Biochim Biophys Acta. 1797(6-7):907-12.

Shao, J., Z. Fu, Y. Ji, X. Guan, S. Guo, Z. Ding, X. Yang, Y. Cong, and Y. Shen. (2016). Leucine zipper-EF-hand containing transmembrane protein 1 (LETM1) forms a Ca2+/H+ antiporter. Sci Rep 6: 34174.

Tsai MF., Jiang D., Zhao L., Clapham D. and Miller C. (2014). Functional reconstitution of the mitochondrial Ca2+/H+ antiporter Letm1. J Gen Physiol. 143(1):67-73.

Waldeck-Weiermair, M., C. Jean-Quartier, R. Rost, M.J. Khan, N. Vishnu, A.I. Bondarenko, H. Imamura, R. Malli, and W.F. Graier. (2011). Leucine zipper EF hand-containing transmembrane protein 1 (Letm1) and uncoupling proteins 2 and 3 (UCP2/3) contribute to two distinct mitochondrial Ca2+ uptake pathways. J. Biol. Chem. 286: 28444-28455.

Zhang, X., G. Chen, Y. Lu, J. Liu, M. Fang, J. Luo, Q. Cao, and X. Wang. (2014). Association of mitochondrial letm1 with epileptic seizures. Cereb Cortex 24: 2533-2540.


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The leucine zipper-EF band transmembrane protein 1, LetM1.  Reduces mitochondrial Ca2+ uptake in response to cytoplasmic Ca2+entry in pancreatic beta cells (Alam et al. 2012).  LetM1 is a cation/Ca2+ transporter, with apparent affinities of cations in the order Ca2+ = Mn2+ > Gd3+ = La3+ > Sr2+ >> Ba2+, Mg2+, K+, Na+ (Shao et al. 2016). The LetM1 turnover rate is only 2 Ca2+/sec with a Km of ~ 25 microM. LetM1 mediates electroneutral 1 Ca2+/2 H+ antiport (Tsai et al. 2013).  However, NCLX (2.A.19.4.4), rather than LetM1, may mediate miltochondrial Ca2+ extrusion (De Marchi et al. 2014).  Letm1 is associated with seizure attacks in Wolf-Hirschhorn syndrome, and its inhibition and mitochondrial dysfunctions contribute to the development of epileptic seizures. An appropriate LetM1 level may be critical for maintaining normal neuronal functions (Zhang et al. 2014).  Glu221 in the mouse orthologue is essential. The protein is a hexamer with a central cavity and exhibits two different conformational states (Shao et al. 2016).


LetM1 of Homo sapiens (O95202)


Mitochondrial distribution and morphology protein Mdm38p (Yol027c)


Mdm38p of Saccharomyces cerevisiae (Q08179)


The mitochondrial LetM1 Ca2+/H+ antiporter (Jiang et al., 2009).


LetM1 of Drosophila melanogaster (P91927)


The yeast Mdm38 (LETM1; YLH47) is a homologue of the human LETM1, the candidate gene for seizures in Wolf-Hirchhorn syndrome. They may be K+/H+ antiporters (Zotova et al., 2010).


LETM1 of Saccharomyces cerevisiae (Q06493)