Sandbox Reserved 1601

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This Sandbox is Reserved from Jan 13 through September 1, 2020 for use in the course CH462 Biochemistry II taught by R. Jeremy Johnson at the Butler University, Indianapolis, USA. This reservation includes Sandbox Reserved 1598 through Sandbox Reserved 1627.

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Mitochondrial Calcium Uniporter (MCU) (Heumann Test Page)

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You may include any references to papers as in: the use of JSmol in Proteopedia ^[1] or to the article describing Jmol ^[2] to the rescue.

1 Overview
2 Structure
- 2.1 Selectivity Filter
- 2.2 Common Mutations
3 Medical Relevance
4 Regulation/Inhibition

Overview

Reference practice ^[3]

Structure

Selectivity Filter

Common Mutations

Medical Relevance

A number of medical conditions all over the body are caused by disruption of the homeostasis of mitochondrial calcium. Diabetes, heart failure, and cancer are just a few members of this broad group of conditions.

Diabetes

In healthy individuals, the 𝛽-cells in the pancreas are responsible for sensing the concentration of glucose in the bloodstream and releasing the appropriate amount of insulin in response. While the mechanism of this activation isn't entirely understood, we can explain a large portion of it in the context of mitochondrial calcium homeostasis. Increased concentration of glucose causes glycolysis in the cell, which increases the amount of ATP. This increase of ATP closes potassium channels in the membrane of the 𝛽-cell which causes depolarization of the membrane. When a certain threshold potential is reached, calcium channels open and create microdomains of calcium below the plasma membrane which allows insulin release by activatin PKC 𝛽-type II. Furthermore, there is a pool of mitochondria in 𝛽-cells near the calcium channels which take in the calcium through the MCU. The mitochondria then create more ATP which sustains and amplifies insulin secretion ^[3].

Any defect in the MCU affects the homeostasis of calcium in the mitochondria. In this case, it can cause insulin secretion to be diminished which can be a causal factor for diabetes I and II.

Heart Failure

Calcium impacts cardiac function in many ways. It is a key modulator of the cardiac functional cycle made up of excitation, contraction (diastole), and relaxation (systole). It also has an impact in cardiac cell death. Mitochondrial calcium contributes to control of oxidative metabolism in excitation-metabolism (EM) coupling which generates the ATP needed for cardiac excitation and contraction in each heartbeat. In sinoatrial nodal cells, an action potential is created by opening of sodium channels to increase the positive charge of the membrane potential. This opens calcium channels (TTCCs and LTCCs) to increase cytosolic calcium levels which activates mitochondrial function and ATP production. This also causes calcium-induced calcium release (CICR) in which the presence of calcium causes the release of more calcium. This initiates muscle contraction by binding troponin C on microfilaments and promotes calcium uptake into the mitochondria. In summary, mitochondrial calcium uptake provides the link between ATP supply and demand during cardiomyocyte contraction. The MCU favors rapid calcium intake which increases heartbeat frequency.^[3]

Ischemia/reperfusion injury (IRI) is caused by the rapid restoration of oxygen to ischemic (oxygen-deficient) tissues. In ischemic conditions, cells undergo anaerobic glycolysis. Because of the cessation of oxidative phosphorylation, the mitochondrial membrane potential is diminished. Additionally, the cytosolic pH is decreased. This drop in pH causes an increase in calcium concentration in the cytoplasm. When oxygen returns, there's a rapid restoration of membrane potential as oxidative phosphorylation resumes. This provides a strong driving force for the entry of calcium into the mitochondria which triggers mitochondrial calcium overload and cell death.^[4]

Therefore, certain issues with the MCU that cause an imbalance in mitochondrial calcium can lead to heart failure. Additionally, even if there is nothing wrong with the MCU, it can have an impact in conditions like IRI. This makes the MCU an interesting target for therapies for both cardiac conditions and many other ailments.

Cancer

Cancer is another condition that can impacted by the MCU, though not much is known about the exact mechanisms. It has mostly been studied in the context of breat and colorectal cancers. Overexpression or overactivation of the MCU complex was shown to promote cancer proliferation. Additionally, the overexpression of MICU1 and MICU2 was shown to decrease mitochondrial calcium levels and prevent apoptosis in cancer cells.^[4] Again, not much is known about the connection between the MCU and cancer cell growth, but the MCU's control over apoptosis and cell growth indicates that mitochondrial calcium regulation is fundamental to cancer cell growth and migration.

Regulation/Inhibition

Uptake of calcium into the mitochondria is pivotal for signalling and bioenergetic processes, but overload of calcium causes release of cytochrome c, overproduction of reactive oxygen species (ROS), swelling of the mitochondria, and opening of the mitochondrial permeability transition pore (mPTP) which all contribute to cell death. Therefore, the MCU has become a target of interest for therapies for certain conditions (like the ones above).^[4] Part of this process is research that looks into inhibitors for the MCU.

Finding an good inhibitor of MCU is no small task. First of all, in the inhibitors that have already been discovered, there is no apparent structure-activity relationship that could predict their inhibitory activity. Additionally, many inhibitors of the MCU are not selective enough for the MCU or have other off-target effects that negatively affect the cell. Among the discovered inhibitors of the MCU are Mitoxantrone and DS16570511 with DS16570511 being the most potent. Furthermore, NecroX-5, KB-R7943, minocycline, and doxycycline have been shown to have inhibitory activity. However, all of these inhibitors are subject to the issues listed before.^[4]

Inorganic salts and coordination complexes have also been shown to inhibit calcium uptake. Specifically, the trivalent lanthanide ions can competitively inhibit the uniporter because of their similar ionic radii and coordination preferences to calcium. In addition, several transition metal coordination complexes (most notably Co, Cr, and Rh) with amine ligands have been shown to inhibit calcium uptake. Again, there is no apparent structure-activity relationship that predicts this behavior.^[4]

The most well-known and commonly used inhibitor of calcium uptake into the mitochondria is ruthenium red (RuRed). RuRed effectively inhibits calcium uptake without affecting mitochondrial respiration or calcium efflux. Additionally, it has been shown to mitigate tissue damage due to IRI and slow cancer cell migration. The issue with RuRed is that its purification has always been a challenging matter.^[4] Interestingly enough, this led to even more developments in the search for an inhibitor. Many scientists had observed that impure RuRed actually had greater inhibition than pure RuRed. One of the common minor impurities of RuRed, Ru360, was found to be the active component of the RuRed mixtures, meaning it responsible for calcium inhibition. Ru360 is now commercially available and has been widely used for the study of calcium-dependent cellular processes and as a therapeutic agent.

Ru360 was a very successful inhibitor,

This is a sample scene created with SAT to by Group, and another to make of the protein. You can make your own scenes on SAT starting from scratch or loading and editing one of these sample scenes.

References

^[3]

^[5]

^[4]

^[6]

^[7]

↑ Hanson, R. M., Prilusky, J., Renjian, Z., Nakane, T. and Sussman, J. L. (2013), JSmol and the Next-Generation Web-Based Representation of 3D Molecular Structure as Applied to Proteopedia. Isr. J. Chem., 53:207-216. doi:http://dx.doi.org/10.1002/ijch.201300024
↑ Herraez A. Biomolecules in the computer: Jmol to the rescue. Biochem Mol Biol Educ. 2006 Jul;34(4):255-61. doi: 10.1002/bmb.2006.494034042644. PMID:21638687 doi:10.1002/bmb.2006.494034042644
↑ ^3.0 ^3.1 ^3.2 ^3.3 Giorgi C, Marchi S, Pinton P. The machineries, regulation and cellular functions of mitochondrial calcium. Nat Rev Mol Cell Biol. 2018 Nov;19(11):713-730. doi: 10.1038/s41580-018-0052-8. PMID:30143745 doi:http://dx.doi.org/10.1038/s41580-018-0052-8
↑ ^4.0 ^4.1 ^4.2 ^4.3 ^4.4 ^4.5 ^4.6 Woods JJ, Wilson JJ. Inhibitors of the mitochondrial calcium uniporter for the treatment of disease. Curr Opin Chem Biol. 2019 Dec 20;55:9-18. doi: 10.1016/j.cbpa.2019.11.006. PMID:31869674 doi:http://dx.doi.org/10.1016/j.cbpa.2019.11.006
↑ Fan C, Fan M, Orlando BJ, Fastman NM, Zhang J, Xu Y, Chambers MG, Xu X, Perry K, Liao M, Feng L. X-ray and cryo-EM structures of the mitochondrial calcium uniporter. Nature. 2018 Jul 11. pii: 10.1038/s41586-018-0330-9. doi:, 10.1038/s41586-018-0330-9. PMID:29995856 doi:http://dx.doi.org/10.1038/s41586-018-0330-9
↑ Kamer KJ, Jiang W, Kaushik VK, Mootha VK, Grabarek Z. Crystal structure of MICU2 and comparison with MICU1 reveal insights into the uniporter gating mechanism. Proc Natl Acad Sci U S A. 2019 Feb 12. pii: 1817759116. doi:, 10.1073/pnas.1817759116. PMID:30755530 doi:http://dx.doi.org/10.1073/pnas.1817759116
↑ Baradaran R, Wang C, Siliciano AF, Long SB. Cryo-EM structures of fungal and metazoan mitochondrial calcium uniporters. Nature. 2018 Jul 11. pii: 10.1038/s41586-018-0331-8. doi:, 10.1038/s41586-018-0331-8. PMID:29995857 doi:http://dx.doi.org/10.1038/s41586-018-0331-8

Retrieved from "http://52.214.119.220/wiki/index.php/Sandbox_Reserved_1601"

@@ Line 38: / Line 38: @@
 == Regulation/Inhibition ==
+Uptake of calcium into the mitochondria is pivotal for signalling and bioenergetic processes, but overload of calcium causes release of cytochrome c, overproduction of reactive oxygen species (ROS), swelling of the mitochondria, and opening of the mitochondrial permeability transition pore (mPTP) which all contribute to cell death.  Therefore, the MCU has become a target of interest for therapies for certain conditions (like the ones above).<ref name="Woods" />  Part of this process is research that looks into inhibitors for the MCU.
+Finding an good inhibitor of MCU is no small task.  First of all, in the inhibitors that have already been discovered, there is no apparent structure-activity relationship that could predict their inhibitory activity.  Additionally, many inhibitors of the MCU are not selective enough for the MCU or have other off-target effects that negatively affect the cell.  Among the discovered inhibitors of the MCU are Mitoxantrone and DS16570511 with DS16570511 being the most potent.  Furthermore, NecroX-5, KB-R7943, minocycline, and doxycycline have been shown to have inhibitory activity.  However, all of these inhibitors are subject to the issues listed before.<ref name="Woods" />
+Inorganic salts and coordination complexes have also been shown to inhibit calcium uptake.  Specifically, the trivalent lanthanide ions can competitively inhibit the uniporter because of their similar ionic radii and coordination preferences to calcium.  In addition, several transition metal coordination complexes (most notably Co, Cr, and Rh) with amine ligands have been shown to inhibit calcium uptake.  Again, there is no apparent structure-activity relationship that predicts this behavior.<ref name="Woods" />
+The most well-known and commonly used inhibitor of calcium uptake into the mitochondria is ruthenium red (RuRed).  RuRed effectively inhibits calcium uptake without affecting mitochondrial respiration or calcium efflux.  Additionally, it has been shown to mitigate tissue damage due to IRI and slow cancer cell migration. The issue with RuRed is that its purification has always been a challenging matter.<ref name="Woods" />  Interestingly enough, this led to even more developments in the search for an inhibitor.  Many scientists had observed that impure RuRed actually had greater inhibition than pure RuRed.  One of the common minor impurities of RuRed, Ru360, was found to be the active component of the RuRed mixtures, meaning it responsible for calcium inhibition. Ru360 is now commercially available and has been widely used for the study of calcium-dependent cellular processes and as a therapeutic agent.
+Ru360 was a very successful inhibitor,
 This is a sample scene created with SAT to <scene name="/12/3456/Sample/1">color</scene> by Group, and another to make <scene name="/12/3456/Sample/2">a transparent representation</scene> of the protein. You can make your own scenes on SAT starting from scratch or loading and editing one of these sample scenes.