Gamma-butyrobetaine dioxygenase

Gamma-butyrobetaine dioxygenase (also known as BBOX, GBBH or γ-butyrobetaine hydroxylase) is an enzyme that in humans is encoded by the BBOX1 gene.[5][6] Gamma-butyrobetaine dioxygenase catalyses the formation of L-carnitine from gamma-butyrobetaine, the last step in the L-carnitine biosynthesis pathway.[7] Carnitine is essential for the transport of activated fatty acids across the mitochondrial membrane during mitochondrial beta oxidation.[6] In humans, gamma-butyrobetaine dioxygenase can be found in the kidney (high), liver (moderate), and brain (very low).[5][8] BBOX1 has recently been identified as a potential cancer gene based on a large-scale microarray data analysis.[9]

BBOX1
Available structures
PDBOrtholog search: PDBe RCSB
Identifiers
AliasesBBOX1, BBH, BBOX, G-BBH, gamma-BBH, gamma-butyrobetaine hydroxylase 1
External IDsOMIM: 603312; MGI: 1891372; HomoloGene: 2967; GeneCards: BBOX1; OMA:BBOX1 - orthologs
Orthologs
SpeciesHumanMouse
Entrez
Ensembl
UniProt
RefSeq (mRNA)

NM_003986
NM_001376258
NM_001376259
NM_001376260
NM_001376261

NM_130452

RefSeq (protein)

NP_003977
NP_001363187
NP_001363188
NP_001363189
NP_001363190

NP_569719

Location (UCSC)Chr 11: 27.04 – 27.13 MbChr 2: 110.09 – 110.14 Mb
PubMed search[3][4]
Wikidata
View/Edit HumanView/Edit Mouse

Reaction

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gamma-butyrobetaine dioxygenase
Identifiers
EC no.1.14.11.1
CAS no.9045-31-2
Databases
IntEnzIntEnz view
BRENDABRENDA entry
ExPASyNiceZyme view
KEGGKEGG entry
MetaCycmetabolic pathway
PRIAMprofile
PDB structuresRCSB PDB PDBe PDBsum
Gene OntologyAmiGO / QuickGO
Search
PMCarticles
PubMedarticles
NCBIproteins

Gamma-butyrobetaine dioxygenase belongs to the 2-oxoglutarate (2OG)-dependent dioxygenase superfamily. It catalyses the following reaction:

4-trimethylammoniobutanoate (γ-butyrobetaine) + 2-oxoglutarate + O2   3-hydroxy-4-trimethylammoniobutanoate (L-carnitine) + succinate + CO2

The three substrates of this enzyme are 4-trimethylammoniobutanoate (γ-butyrobetaine), 2-oxoglutarate, and O2,[10] whereas its three products are 3-hydroxy-4-trimethylammoniobutanoate (L-carnitine), succinate, and carbon dioxide.

This enzyme belongs to the family of oxidoreductases, specifically those acting on paired donors, with O2 as oxidant and incorporation or reduction of oxygen. The oxygen incorporated need not be derived from O2 with 2-oxoglutarate as one donor, and incorporation of one atom of oxygen into each donor. This enzyme participates in lysine degradation. Iron is a cofactor for gamma-butyrobetaine dioxygenase. Similar to many other 2OG oxygenases, the activity of gamma-butyrobetaine dioxygenase can be stimulated by reducing agents such as ascorbate and glutathione.[11][12][13][14] The catalytic activity of gamma-butyrobetaine dioxygenase can be stimulated with different metal ions, especially potassium ions.[15]

Both the apo (PDB id: 3N6W)[16] and the holo (PDB id: 3O2G)[17] structures of gamma-butyrobetaine dioxygenase have been solved, demonstrating an induced fit mechanism may contribute to the catalytic activity of gamma-butyrobetaine dioxygenase.

Gamma-butyrobetaine dioxygenase is promiscuous in substrate selectivity and it processes a number of modified substrates, including the natural catalytic products L-carnitine and D-carnitine, forming 3-dehydrocarnitine and trimethylaminoacetone.[17][18] Gamma-butyrobetaine dioxygenase also catalyses the oxidation of mildronate[19] to form multiple products including malonic acid semialdehyde, dimethylamine, formaldehyde and (1-methylimidazolidin-4-yl)acetic acid, which is proposed to be formed via a Stevens rearrangement mechanism.[20][21] Gamma-butyrobetaine dioxygenase is unique among other human 2OG oxygenases that it catalyses both hydroxylation (e.g.: L-carnitine), demethylation (e.g.: formaldehyde) and C-C bond formation (e.g.: (1-methylimidazolidin-4-yl)acetic acid).[22]

Inhibition

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Gamma-butyrobetaine dioxygenase is an inhibition target for 3-(2,2,2-trimethylhydraziniumyl)propionate (mildronate, also known as THP, MET-88, Meldonium or Quarterine). Mildronate is offered, clinically, to non-U.S. markets, in treatment of angina and myocardial infarction.[23][24][25] Some studies suggested that mildronate may also be beneficial for the treatment of neurological disorder,[26][27] diabetes,[28] and seizures and alcohol intoxication.[29] Mildronate is currently manufactured and marketed by Grindeks, a pharmaceutical company based in Latvia. To date, at least five clinical trial reports were published in peer-reviewed journals documenting the efficacy and safety of mildronate on the treatments of angina, stroke and chronic heart failure.[30][31][32][33][34] However, there have been no randomized clinical trials to support the use of mildronate to treat any cardiovascular disease.[35][better source needed]

Mildronate has a similar structure to the natural substrate gamma-butyrobetaine, with a NH group replacing the CH2 of gamma-butyrobetaine at the C-4 position. A crystal structure of mldronate in complex with gamma-butyrobetaine dioxygenase was published, and it suggests mildronate bind to gamma-butyrobetaine dioxygenase in exactly the same way as gamma-butyrobetaine (PDB id: 3MS5).[36] To date, most enzyme inhibitors for human 2OG oxygenases bind to the cosubstrate 2OG binding site; mildronate is a rare example of a non-peptidyl substrate mimic inhibitor.[37] Although initial reports suggested mildronate is a non-competitive and non-hydroxylatable analogue of gamma-butyrobetaine,[38] further studies have identified mildronate is indeed a substrate for gamma-butyrobetaine dioxygenase.[17][20][39]

Similar to other 2OG oxygenases, gamma-butyrobetaine dioxygenase can be inhibited by 2OG mimics and aromatic inhibitors such as pyridine 2,4-dicarboxylate.[40] Other reported gamma-butyrobetaine dioxygenase inhibitors include cyclopropyl-substituted gamma-butyrobetaines[41] and 3-(2,2-dimethylcyclopropyl)propanoic acid, which is a mechanism-based enzyme inhibitor.[42]

Assay

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Several in vitro biochemical assays have been applied to monitor the catalytic activity of gamma-butyrobetaine dioxygenase. Early methods have mainly focused on the use of radiolabeled compounds, including 14C-labelled gamma-butyrobetaine[43] and 14C-labelled 2OG.[44] Enzyme-coupled method have also been applied to detect carnitine formation, by using the enzyme carnitine acetyltransferase and 14C-labelled acetyl-coenzyme A to give labelled acetylcarnitine for detection. Using this method, it is possible to detect carnitine concentration down to the pico-molar range.[45][46][47] Other analytical methods including mass spectrometry and NMR have also been applied,[17] and they are in particularly useful for the study of the coupling ratio between 2OG oxidation and substrate formation, and for the characterisation of unknown enzymatic products.[18] However, these methods are often not suitable for high-throughput screening and require expensive instrumentation. A potentially high-throughput fluorescence-based assay has also been proposed by using a fluorinated-gamma-butyrobetaine analog.[48] The fluoride ions released as a result of gamma-butyrobetaine dioxygenase catalyses can be detected by using chemosensors such as protected fluorescein.[49]

See also

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References

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  1. ^ a b c GRCh38: Ensembl release 89: ENSG00000129151Ensembl, May 2017
  2. ^ a b c GRCm38: Ensembl release 89: ENSMUSG00000041660Ensembl, May 2017
  3. ^ "Human PubMed Reference:". National Center for Biotechnology Information, U.S. National Library of Medicine.
  4. ^ "Mouse PubMed Reference:". National Center for Biotechnology Information, U.S. National Library of Medicine.
  5. ^ a b Vaz FM, van Gool S, Ofman R, Ijlst L, Wanders RJ (Sep 1998). "Carnitine biosynthesis: identification of the cDNA encoding human gamma-butyrobetaine hydroxylase". Biochemical and Biophysical Research Communications. 250 (2): 506–10. doi:10.1006/bbrc.1998.9343. PMID 9753662.
  6. ^ a b "Entrez Gene: BBOX1 butyrobetaine (gamma), 2-oxoglutarate dioxygenase (gamma-butyrobetaine hydroxylase) 1".
  7. ^ Paul HS, Sekas G, Adibi SA (Feb 1992). "Carnitine biosynthesis in hepatic peroxisomes. Demonstration of gamma-butyrobetaine hydroxylase activity". European Journal of Biochemistry. 203 (3): 599–605. doi:10.1111/j.1432-1033.1992.tb16589.x. PMID 1735445.
  8. ^ Lindstedt G, Lindstedt S, Nordin I (Oct 1982). "Gamma-butyrobetaine hydroxylase in human kidney". Scandinavian Journal of Clinical and Laboratory Investigation. 42 (6): 477–85. doi:10.3109/00365518209168117. PMID 7156861.
  9. ^ Dawany NB, Dampier WN, Tozeren A (Jun 2011). "Large-scale integration of microarray data reveals genes and pathways common to multiple cancer types". International Journal of Cancer. 128 (12): 2881–91. doi:10.1002/ijc.25854. PMID 21165954. S2CID 24740881.
  10. ^ Lindstedt G, Lindstedt S (Aug 1970). "Cofactor requirements of gamma-butyrobetaine hydroxylase from rat liver". The Journal of Biological Chemistry. 245 (16): 4178–86. doi:10.1016/S0021-9258(18)62901-1. PMID 4396068.
  11. ^ Rebouche CJ (Dec 1991). "Ascorbic acid and carnitine biosynthesis". The American Journal of Clinical Nutrition. 54 (6 Suppl): 1147S–1152S. doi:10.1093/ajcn/54.6.1147s. PMID 1962562.
  12. ^ Nelson PJ, Pruitt RE, Henderson LL, Jenness R, Henderson LM (Jan 1981). "Effect of ascorbic acid deficiency on the in vivo synthesis of carnitine". Biochimica et Biophysica Acta (BBA) - General Subjects. 672 (1): 123–7. doi:10.1016/0304-4165(81)90286-5. PMID 6783120.
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  16. ^ PDB: 3N6W​;Tars K, Rumnieks J, Zeltins A, Kazaks A, Kotelovica S, Leonciks A, Sharipo J, Viksna A, Kuka J, Liepinsh E, Dambrova M (Aug 2010). "Crystal structure of human gamma-butyrobetaine hydroxylase". Biochemical and Biophysical Research Communications. 398 (4): 634–9. doi:10.1016/j.bbrc.2010.06.121. PMID 20599753.
  17. ^ a b c d PDB: 3O2G​; Leung IK, Krojer TJ, Kochan GT, Henry L, von Delft F, Claridge TD, Oppermann U, McDonough MA, Schofield CJ (Dec 2010). "Structural and mechanistic studies on γ-butyrobetaine hydroxylase". Chemistry & Biology. 17 (12): 1316–24. doi:10.1016/j.chembiol.2010.09.016. PMID 21168767.
  18. ^ a b Fujimori DG (Dec 2010). "A novel enzymatic rearrangement". Chemistry & Biology. 17 (12): 1269–70. doi:10.1016/j.chembiol.2010.12.003. PMID 21168760.
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  25. ^ Hayashi Y, Kirimoto T, Asaka N, Nakano M, Tajima K, Miyake H, Matsuura N (May 2000). "Beneficial effects of MET-88, a gamma-butyrobetaine hydroxylase inhibitor in rats with heart failure following myocardial infarction". European Journal of Pharmacology. 395 (3): 217–24. doi:10.1016/S0014-2999(00)00098-4. PMID 10812052.
  26. ^ Sjakste N, Gutcaits A, Kalvinsh I (2005). "Mildronate: an antiischemic drug for neurological indications". CNS Drug Reviews. 11 (2): 151–68. doi:10.1111/j.1527-3458.2005.tb00267.x. PMC 6741751. PMID 16007237.
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  28. ^ Liepinsh E, Skapare E, Svalbe B, Makrecka M, Cirule H, Dambrova M (May 2011). "Anti-diabetic effects of mildronate alone or in combination with metformin in obese Zucker rats". European Journal of Pharmacology. 658 (2–3): 277–83. doi:10.1016/j.ejphar.2011.02.019. PMID 21371472.
  29. ^ Zvejniece L, Svalbe B, Makrecka M, Liepinsh E, Kalvinsh I, Dambrova M (Sep 2010). "Mildronate exerts acute anticonvulsant and antihypnotic effects". Behavioural Pharmacology. 21 (5–6): 548–55. doi:10.1097/FBP.0b013e32833d5a59. PMID 20661137. S2CID 12501700.
  30. ^ Dzerve V, Matisone D, Kukulis I, Romanova J, Putane L, Grabauskiene V, Skarda I, Berzina D, Strautmanis J (2005). "Mildronate improves peripheral circulation in patients with chronic heart failure: results of a clinical trial (the first report)" (PDF). Semin Cardiol. 11 (2): 56–64. ISSN 1648-7966. Archived from the original (PDF) on 2012-03-28.
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  36. ^ PDB: 3MS5
  37. ^ Rose NR, McDonough MA, King ON, Kawamura A, Schofield CJ (Aug 2011). "Inhibition of 2-oxoglutarate dependent oxygenases". Chemical Society Reviews. 40 (8): 4364–97. doi:10.1039/c0cs00203h. PMID 21390379.
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  40. ^ Ng SF, Hanauske-Abel HM, Englard S (Jan 1991). "Cosubstrate binding site of Pseudomonas sp. AK1 gamma-butyrobetaine hydroxylase. Interactions with structural analogs of alpha-ketoglutarate". The Journal of Biological Chemistry. 266 (3): 1526–33. doi:10.1016/S0021-9258(18)52326-7. PMID 1988434.
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  48. ^ Rydzik AM, Leung IK, Kochan GT, Thalhammer A, Oppermann U, Claridge TD, Schofield CJ (Jul 2012). "Development and application of a fluoride-detection-based fluorescence assay for γ-butyrobetaine hydroxylase". ChemBioChem. 13 (11): 1559–63. doi:10.1002/cbic.201200256. PMID 22730246. S2CID 13956474.
  49. ^ Cametti M, Rissanen K (May 2009). "Recognition and sensing of fluoride anion". Chemical Communications (20): 2809–29. doi:10.1039/B902069A. PMID 19436879.

Further reading

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