Stereochemistry of the Reactions of Glutamate-1-semialdehyde Aminomutase with 4,5-Diaminovalerate

Simona D'Aguanno, Isabel Nogues Gonzales, Maurizio Simmaco, Roberto Contestabile, Robert A. John

Research output: Contribution to journalArticle

Abstract

Conversion of glutamate 1-semialdehyde to the tetrapyrrole precursor, 5-aminolevulinate, takes place in an aminomutase-catalyzed reaction involving transformations at both the non-chiral C5 and the chiral C4 of the intermediate 4,5-diaminovalerate. Presented with racemic diaminovalerate and an excess of succinic semialdehyde, the enzyme catalyzes a transamination in which only the L-enantiomer is consumed. Simultaneously, equimolar 4-aminobutyrate and aminolevulinate are formed. The enzyme is also shown to transaminate aminolevulinate and 4-aminohexenoate to L-diaminovalerate as the exclusive amino product. The interaction of the enzyme with pure D- and L-enantiomers of diaminovalerate prepared by these reactions is described. Transamination of L-diaminovalerate yielded aminolevulinate quantitatively showing that reaction at the C5 amine does not occur significantly. A much slower transamination reaction was catalyzed with D-diaminovalerate as substrate. One product of this reaction, 4-aminobutyrate, was formed in the amount equal to that of the diaminovalerate consumed. Glutamate semialdehyde was deduced to be the other primary product and was also measured in significant amounts when a high concentration of the enzyme in its pyridoxal form was reacted with D-diaminovalerate in a single turnover. Single turnover reactions showed that both enantiomers of diaminovalerate converted the enzyme from its 420-nm absorbing pyridoxaldimine form to the 330-nm absorbing pyridoxamine via rapidly formed intermediates with different absorption spectra. The intermediate formed with L-DAVA (λ max = 420 nm) was deduced to be the protonated external aldimine with the 4-amino group. The intermediate formed with D-DAVA (λ max = 390 nm) was deduced to be the unprotonated external aldimine with the 5-amino group.

Original languageEnglish
Pages (from-to)40521-40526
Number of pages6
JournalJournal of Biological Chemistry
Volume278
Issue number42
DOIs
Publication statusPublished - Oct 17 2003

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Stereochemistry
Enantiomers
Aminobutyrates
Enzymes
Pyridoxamine
Tetrapyrroles
Pyridoxal
Aminolevulinic Acid
Amines
Absorption spectra
Glutamic Acid
glutamate-1-semialdehyde
Substrates

ASJC Scopus subject areas

  • Biochemistry

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Stereochemistry of the Reactions of Glutamate-1-semialdehyde Aminomutase with 4,5-Diaminovalerate. / D'Aguanno, Simona; Gonzales, Isabel Nogues; Simmaco, Maurizio; Contestabile, Roberto; John, Robert A.

In: Journal of Biological Chemistry, Vol. 278, No. 42, 17.10.2003, p. 40521-40526.

Research output: Contribution to journalArticle

D'Aguanno, Simona ; Gonzales, Isabel Nogues ; Simmaco, Maurizio ; Contestabile, Roberto ; John, Robert A. / Stereochemistry of the Reactions of Glutamate-1-semialdehyde Aminomutase with 4,5-Diaminovalerate. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 42. pp. 40521-40526.
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abstract = "Conversion of glutamate 1-semialdehyde to the tetrapyrrole precursor, 5-aminolevulinate, takes place in an aminomutase-catalyzed reaction involving transformations at both the non-chiral C5 and the chiral C4 of the intermediate 4,5-diaminovalerate. Presented with racemic diaminovalerate and an excess of succinic semialdehyde, the enzyme catalyzes a transamination in which only the L-enantiomer is consumed. Simultaneously, equimolar 4-aminobutyrate and aminolevulinate are formed. The enzyme is also shown to transaminate aminolevulinate and 4-aminohexenoate to L-diaminovalerate as the exclusive amino product. The interaction of the enzyme with pure D- and L-enantiomers of diaminovalerate prepared by these reactions is described. Transamination of L-diaminovalerate yielded aminolevulinate quantitatively showing that reaction at the C5 amine does not occur significantly. A much slower transamination reaction was catalyzed with D-diaminovalerate as substrate. One product of this reaction, 4-aminobutyrate, was formed in the amount equal to that of the diaminovalerate consumed. Glutamate semialdehyde was deduced to be the other primary product and was also measured in significant amounts when a high concentration of the enzyme in its pyridoxal form was reacted with D-diaminovalerate in a single turnover. Single turnover reactions showed that both enantiomers of diaminovalerate converted the enzyme from its 420-nm absorbing pyridoxaldimine form to the 330-nm absorbing pyridoxamine via rapidly formed intermediates with different absorption spectra. The intermediate formed with L-DAVA (λ max = 420 nm) was deduced to be the protonated external aldimine with the 4-amino group. The intermediate formed with D-DAVA (λ max = 390 nm) was deduced to be the unprotonated external aldimine with the 5-amino group.",
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AB - Conversion of glutamate 1-semialdehyde to the tetrapyrrole precursor, 5-aminolevulinate, takes place in an aminomutase-catalyzed reaction involving transformations at both the non-chiral C5 and the chiral C4 of the intermediate 4,5-diaminovalerate. Presented with racemic diaminovalerate and an excess of succinic semialdehyde, the enzyme catalyzes a transamination in which only the L-enantiomer is consumed. Simultaneously, equimolar 4-aminobutyrate and aminolevulinate are formed. The enzyme is also shown to transaminate aminolevulinate and 4-aminohexenoate to L-diaminovalerate as the exclusive amino product. The interaction of the enzyme with pure D- and L-enantiomers of diaminovalerate prepared by these reactions is described. Transamination of L-diaminovalerate yielded aminolevulinate quantitatively showing that reaction at the C5 amine does not occur significantly. A much slower transamination reaction was catalyzed with D-diaminovalerate as substrate. One product of this reaction, 4-aminobutyrate, was formed in the amount equal to that of the diaminovalerate consumed. Glutamate semialdehyde was deduced to be the other primary product and was also measured in significant amounts when a high concentration of the enzyme in its pyridoxal form was reacted with D-diaminovalerate in a single turnover. Single turnover reactions showed that both enantiomers of diaminovalerate converted the enzyme from its 420-nm absorbing pyridoxaldimine form to the 330-nm absorbing pyridoxamine via rapidly formed intermediates with different absorption spectra. The intermediate formed with L-DAVA (λ max = 420 nm) was deduced to be the protonated external aldimine with the 4-amino group. The intermediate formed with D-DAVA (λ max = 390 nm) was deduced to be the unprotonated external aldimine with the 5-amino group.

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