2-isopropylmalate synthase

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2-isopropylmalate synthase
Identifiers
EC no. 2.3.3.13
CAS no. 9030-98-2
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MetaCyc metabolic pathway
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In enzymology, a 2-isopropylmalate synthase (EC 2.3.3.13) is an enzyme that catalyzes the chemical reaction

acetyl-CoA + 3-methyl-2-oxobutanoate + H2O (2S)-2-isopropylmalate + CoA

The three substrates of this enzyme are acetyl-CoA, 3-methyl-2-oxobutanoate, and H2O, and its products are (2S)-2-isopropylmalate and CoA.

The enzyme belongs to the family of transferases, specifically those acyltransferases that convert acyl groups into alkyl groups on transfer. The systematic name of this enzyme class is acetyl-CoA:3-methyl-2-oxobutanoate C-acetyltransferase (thioester-hydrolysing, carboxymethyl-forming). Other names in common use include 3-carboxy-3-hydroxy-4-methylpentanoate 3-methyl-2-oxobutanoate-lyase, (CoA-acetylating), alpha-isopropylmalate synthetase, alpha-isopropylmalate synthase, alpha-isopropylmalic synthetase, isopropylmalate synthase, and isopropylmalate synthetase. This enzyme participates in biosynthesis of L-leucine and pyruvate metabolism. Monovalent and divalent cation activation have been reported for enzymes from different sources. [1] [2] [3]

Mycobacterium tuberculosis α-isopropylmalate synthase requires a divalent metal ion, of which Mg2+ and Mn2+ give highest activity, and a monovalent cation, with K+ as the best activator. [4] [5] Zn2+ was shown to be an inhibitor, contrary to what was assumed from the structural data. Another feature of the M. tuberculosis homolog is that L-leucine, the feedback inhibitor, inhibits the enzyme in a time-dependent fashion. This was the first demonstration of a feedback inhibitor that displays slow-onset inhibition. [6]

Tertiary structure

As of late 2007, only one tertiary structure has been solved for this class of enzymes, with the Protein Data Bank accession code 1SR9.

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References

  1. Cole FE, Kalyanpur MG, Stevens CM (1973). "Absolute configuration of alpha isopropylmalate and the mechanism of its conversion to beta isopropylmalate in the biosynthesis of leucine". Biochemistry. 12 (17): 3346–50. doi:10.1021/bi00741a031. PMID   4270046.
  2. Kohlhaw G, Leary TR, Umbarger HE (1969). "Alpha-isopropylmalate synthase from Salmonella typhimurium Purification and properties". J. Biol. Chem. 244 (8): 2218–25. doi: 10.1016/S0021-9258(18)97789-6 . PMID   4976555.
  3. Webster RE; Gross, SR (1965). "The alpha-isopropylmalate synthetase of Neurospora. I. The kinetics and end product control of alpha-isopropylmalate synthetase function". Biochemistry. 4 (11): 2309–2327. doi:10.1021/bi00887a008.
  4. Carvalho LP, Blanchard, JS (2006). "Kinetic and Chemical Mechanism of alpha-Isopropylmalate Synthase from Mycobacterium tuberculosis". Biochemistry. 45 (29): 8988–99. doi:10.1021/bi0606602. PMC   2507874 . PMID   16846242.
  5. Carvalho LP, Blanchard, JS (2006). "Kinetic analysis of the effects of monovalent cations and divalent metals on the activity of Mycobacterium tuberculosis alpha-isopropylmalate synthase". Archives of Biochemistry and Biophysics. 451 (2): 141–48. doi:10.1016/j.abb.2006.03.030. PMID   16684501.
  6. Carvalho LP, Argyrou A, Blanchard, JS (2005). "Slow-onset Feedback Inhibition: Inhibition of Mycobacterium tuberculosis alpha-Isopropylmalate Synthase by L-Leucine". Journal of the American Chemical Society. 127 (28): 10004–5. doi:10.1021/ja052513h. PMID   16011356.