Branched-chain alpha-keto acid dehydrogenase complex

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The branched-chain α-ketoacid dehydrogenase complex (BCKDC) is a multi-subunit complex of enzymes that is found on the mitochondrial inner membrane.[1] This enzyme complex catalyzes the oxidative decarboxylation of branched, short-chain alpha-ketoacids. BCKDC is a member of the mitochondrial α-ketoacid dehydrogenase complex family comprising pyruvate dehydrogenase and alpha-ketoglutarate dehydrogenase, key enzymes that function in the Krebs cycle.

Coenzymes

This complex requires the following 5 coenzymes:

Biological Function

In animal tissue, BCKDC catalyzes an irreversible step[2] in the catabolism of branched-chain amino acids—namely L-isoleucine, L-valine, L-leucine L-threonine and their derivatives (L-alpha-keto-beta-methylvalerate, alpha-ketoisovalerate, and alpha-ketoisocaproate, and alpha-ketobutyrate respectively).[3][4][5] In bacteria, this enzyme participates in the synthesis of branched, long-chain fatty acids.[6] In plants, this enzyme is involved in the synthesis of branched, long-chain hydrocarbons.

The overall catabolic reaction catalyzed by the BCKDC is shown in Figure 1.

File:Overall reaction catalyzed by the branched-chain alpha-ketoacid dehydrogenase complex.png
Figure 1: This is the overall reaction catalyzed by the branched-chain alpha-ketoacid dehydrogenase complex.

Structure

The mechanism of enzymatic catalysis by the BCKDC draws largely upon the elaborate structure of this large enzyme complex. This enzyme complex is composed of three catalytic components: alpha-ketoacid dehydrogenase (also referred to as the E1 component), dihydrolipoyl transacylase (E2 component), and dihydrolipoamide dehydrogenase (E3 component). In humans, 24 copies of E2 arranged in octahedral symmetry form the core of the BCKDC.[7] Non-covalently linked to this polymer of 24 E2 subunits are 12 E1 α2β2 tetramers and 6 E3 homodimers. In addition to the E1/E3-binding domain, there are 2 other important structural domains in the E2 subunit: (i) a lipoyl-bearing domain in the amino-terminal portion of the protein and (ii) an inner-core domain in the carboxy-terminal portion. The inner-core domain is linked to the other two domains of the E2 subunit by two interdomain segments (linkers).[8] The inner-core domain is necessary to form the oligomeric core of the enzyme complex and catalyzes the acyltransferase reaction (shown in the "Mechanism" section below).[9] The lipoyl domain of E2 is free to swing between the active sites of the E1, E2, and E3 subunits on the assembled BCKDC by virtue of the conformational flexibility of the aforementioned linkers (see Figure 2).[10][11] Thus, in terms of function as well as structure, the E2 component plays a central role in the overall reaction catalyzed by the BCKDC.

File:Swinging arm of BCKDC.png
Figure 2:This is a schematic of the "swinging" lipoyl domain. Note that this lipoyl domain is covalently attached to the E2 subunit of the BCKDC, but is free to swing between the E1, E2, and E3 subunits. As is described in the "Mechanism" section, the ability of the lipoyl group to freely swing between the active sites on each of the three subunits of the BCKDC plays a large and important role in the catalytic activity of this enzyme complex.[12]

The role of each subunit is as follows:

E1 subunit

E1 uses thiamine pyrophosphate (TPP) as a catalytic cofactor. E1 catalyzes both the decarboxylation of the α-ketoacid and the subsequent reductive acylation of the lipoyl moiety (another catalytic cofactor) that is covalently bound to E2.

E2 subunit

E2 catalyzes a transfer of the acyl group from the lipoyl moiety to coenzyme A (a stoichiometric cofactor).[13]

E3 subunit

The E3 component is a flavoprotein, and it re-oxidizes the reduced lipoyl sulfur residues of E2 using FAD (a catalytic cofactor) as the oxidant. FAD then transfers these protons and electrons to NAD+ (a stoichiometric cofactor) to complete the reaction cycle.

Mechanism

As previously mentioned, BCKDC’s primary function in mammals is to catalyze an irreversible step in the catabolism of branched-chain amino acids. However, the BCKDC has a relatively broad specificity, also oxidizing 4-methylthio-2-oxobutyrate and 2-oxobutyrate at comparable rates and with similar Km values as for its branched-chain amino acid substrates.[14] The BCKDC will also oxidize pyruvate, but at such a slow rate this side reaction has very little physiological significance.[15][16]

The reaction mechanism is as follows.[17] Please note that any of several branched-chain α-ketoacids could have been used as a starting material; for this example, α-ketoisovalerate was arbitrarily chosen as the BCKDC substrate.

NOTE: Steps 1 and 2 occur in the E1 domain

STEP 1: α-ketoisovalerate combines with TPP and is then decarboxylated. The proper arrow-pushing mechanism is shown in Figure 3.

File:BCKDC catalytic mechanism step 1.png
Figure 3: α-ketoisovalerate combines with TPP and is then decarboxylated

STEP 2: The 2-methylpropanol-TPP is oxidized to form an acyl group while being simultaneously transferred to the lipoyl cofactor on E2. Note that TPP is regenerated. The proper arrow-pushing mechanism is shown in Figure 4.

File:BCKDC catalytic mechanism step 2.png
Figure 4: The 2-methylpropanol-TPP is oxidized to form an acyl group while being simultaneously transferred to the lipoyl cofactor on E2. Note that TPP is regenerated.
NOTE: The acylated lipoyl arm now leaves E1 and swings into the E2 active site, where Step 3 occurs.

STEP 3: Acyl group transfer to CoA. The proper arrow-pushing mechanism is shown in Figure 5.

File:BCKDC catalytic mechanism step 3.png
Figure 5: Acyl group transfer to CoA
*NOTE: The reduced lipoyl arm now swings into the E3 active site, where Steps 4 and 5 occur.

STEP 4: Oxidation of the lipoyl moiety by the FAD coenzyme, as shown in Figure 6.

File:BCKDC catalytic mechanism step 4.png
Figure 6: Oxidation of the lipoyl moiety by the FAD coenzyme.

STEP 5: Reoxidation of FADH2 to FAD, producing NADH:

FADH2 + NAD+ --> FAD + NADH + H+

Disease Relevance

A deficiency in any of the enzymes of this complex as well as an inhibition of the complex as a whole leads to a buildup of branched-chain amino acids and their harmful derivatives in the body. These accumulations lend a sweet smell to bodily excretions (such as ear wax and urine), leading to a pathology known as maple syrup urine disease.[18]

This enzyme is an autoantigen recognized in primary biliary cirrhosis, a form of acute liver failure. These antibodies appear to recognize oxidized protein that has resulted from inflammatory immune responses. Some of these inflammatory responses are explained by gluten sensitivity.[19] Other mitochondrial autoantigens include pyruvate dehydrogenase and branched-chain oxoglutarate dehydrogenase, which are antigens recognized by anti-mitochondrial antibodies.

References

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  8. ↑ Lua error in package.lua at line 80: module 'strict' not found.
  9. ↑ Lua error in package.lua at line 80: module 'strict' not found.
  10. ↑ Lua error in package.lua at line 80: module 'strict' not found.
  11. ↑ Lua error in package.lua at line 80: module 'strict' not found.
  12. ↑ Berg, Jeremy M., John L. Tymoczko, Lubert Stryer, and Lubert Stryer. Biochemistry. 6th ed. New York: W.H. Freeman, 2007. 481. Print.
  13. ↑ Lua error in package.lua at line 80: module 'strict' not found.
  14. ↑ Lua error in package.lua at line 80: module 'strict' not found.
  15. ↑ Lua error in package.lua at line 80: module 'strict' not found.
  16. ↑ Lua error in package.lua at line 80: module 'strict' not found.
  17. ↑ Berg, Jeremy M., John L. Tymoczko, Lubert Stryer, and Lubert Stryer. Biochemistry. 6th ed. New York: W.H. Freeman, 2007. 478-79. Print.
  18. ↑ Lua error in package.lua at line 80: module 'strict' not found.
  19. ↑ Lua error in package.lua at line 80: module 'strict' not found.

External links

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