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Mitochondrial Calcium Uniporter

spinqualitylabelsall models Calcium Uniporter 6DT0 Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Introduction Calcium is a key signaling molecule involved in many physiological functions including muscle contraction, neuron excitability, cell migration and growth. The mitochondria are important regulators of calcium in the body; they orchestrate the regulation of ATP production, cell death, and intracellular calcium signaling. The process of calcium regulation in the mitochondria is as follows: Calcium moves in one direction through the mitochondria from the intermembrane space through the inner mitochondrial membrane into the matrix. The matrix is more negatively charged driven by the electron transport chain which facilitates calcium movement with its concentration gradient. Maintaining this concentration gradient and the homeostasis of calcium in the mitochondria is the calcium uniporter (MCU). The MCU is a complex composed of regulatory subunits including mitochondrial calcium uptake (MICU), essential MICU regulator (EMRE), MCU regulatory subunit b (MCUb), and MCU regulator 1 (MCUR1). On the outside of the uniporter portion of the MCU are mitochondrial calcium uptake 1 and 2 (MICU1 and MICU2). MICU1 and MICU2 act as gatekeepers by setting the calcium uptake threshold for activation of MCU, only allowing calcium uptake at high calcium concentrations. MICU1 and MICU2 bind together and associate with another external subunit, EMRE, to regulate calcium acquisition by the MCU. EMRE connects the MICU1 and MICU2 sensors to MCU therefore regulating calcium uptake for the protein. Cryogenic electron microscopy (Cryo-EM) was instrumental in outlining the complete structure of this protein. This analysis led to the discovery of key residues within the structure of the MCU as well as providing a structural framework for understanding the mechanism by with the MCU functions. Structural highlights and mechanism The MCU is a dimer of dimers, described as tetrameric truncated pyramid. The uniporter has only a single strong binding site located in the selectivity pore with specificity for Calcium, near the surface of the inner mitochondrial membrane. The Calcium from the cytoplasm enters the mitochondrial inner membrane space through the mitochondrial membrane and is passed to the mitochondrial matrix via the MCU (Figure 1). The transmembrane domain opens to the surface of the inner mitochondrial membrane, while the soluble domain, the coiled coil and the N-terminal domain reside inside the inner mitochondrial membrane, opening to the mitochondrial matrix. Figure 1: Structure of mitochondrial calcium uniporter colored by functional domain designed in PyMol. The transmembrane domain is highlighted salmon, the matrix in light cyan, coiled coil in dark violet, and the N-Terminal Domain in slate blue. [1] 6DT0 Each domain has a different functional role Transmembrane Domain The transmembrane domain is on the inner mitochondrial membrane open to the inner membrane space. The small pore, highly specific for calcium binding is located in transmembrane 2 (TM2) while transmembrane 1 (TM1) surrounds the pore. The transmembrane domain exhibits four fold rotational symmetry. Selectivity Filter The selectivity filter contains Glu358, Trp354, and Pro359 to allow calcium to pass through the uniporter. The carboxylate oxygen of the Glu358 side chains draw in the positive calcium ion. The diameter of the carboxyl ring is about 4Å, allowing only a dehydrated Ca2+ ion to bind. Trp38, which is directly next to the Glu residues, stabilizes the carbonyl side chains through hydrogen bonding and anion pi interactions. These Trp residues also form stacking interactions with Pro359, which orientate the Glu carboxyl side chains towards the middle of the pore to interact with Ca2+ ions. Approximately one helical turn below the glutamate ring of the selectivity filter, there is a tyrosine ring coming a 12Å wide pore allowing high conductivity. The wider opening allows calcium to rehydrate once they pass the selectivity pore. Calcium Uniporter Structure Soluble Domain The coiled coil is the first subsection of the soluble domain, which resides in the inner mitochondrial membrane. The coiled coil functions as the joints of the uniporter, providing flexibility to promote transport of Calcium ions down their concentration gradient. The junction between the transmembrane domain and the coiled coil's flexibility can be attributed to the disordered packing between subunits; subunits A and C adopt different conformations than the B and D subunits, although they superimpose well. Figure 2: Symmetry and organization of subunits from looking down into the uniporter from the inner mitochondrial membrane[2] 6DT0 When calcium binds to the selectivity pore, the coiled coil swings approximately 8° around its end near the N-Terminal Domain. This movement propagates to the top of the transmembrane domain, where the pore is located, about 85 amperes away. The largest displacement triggered by the movement of the coiled coil is in the transmembrane domain, where the coil bends 20°, moving the transmembrane domain further apart. The N-Terminal domain (NTD) is involved in calcium condition. Reorganization in the NTD due to shifts in the coiled coil switch subunits to alter membrane packing causing the interactions between the tyrosines and transmembrane helices. This propagation facilitates a rotamer switch between one pair of tyrosine controlling calcium flow through the pore. The soluble domain is wider than the transmembrane domain, allowing calcium ions to rehydrate and increasing the conductivity of ions through the uniporter into the mitochondrial matrix. Disease Links Types 1 and 2 Diabetes Pancreatic beta cells circulate insulin through the body. Glucose initiates signals allowing these cells to break down sugar and release insulin, which is all stimulated by mitochondrial energy metabolism. Calcium homeostasis plays a fundamental role in ATP production supplying energy to this process. Defects in homeostasis of calcium like chronic calcium depletion, caused by leaky Ryanodine receptors, causes types 1 and 2 diabetes through failure of this mechanism. Treatment involves targeting the MCU and MICU1 to open the calcium channel and allow more uptake of calcium ions into the mitochondria. Heart Failure Calcium overload in the mitochondria of cardiac cells lead to apoptotic cardiac cell death. Calcium governs excitation contraction coupling (EC coupling) of the cardiac muscles, which creates the ATP needed to power the contraction during heart beats. The increase in mitochondrial Ca2+ concentration is essential for the functioning of this muscle contraction. Mitochondrial Ca2+ overload, though, leads to necrotic cardiac cell death and can be targeted with regulation of the MCU. An example of treatment would be with the use of Ru360 to inhibit the uptake of Ca2+ ions into the mitochondria. Student Contributors Holly Rowe Lizzy Ratz Madi Summers