Stable-isotope labeled metabolites of the phytohormone, indole-3-acetic acid

Abstract

1,3‐Dicyclohexylcarbodiimide‐mediated condensation of [3a,4,5,6,7,7a‐13C6]indole‐3‐acetic acid with the bis(tert‐butyl) esters of L‐aspartic or L‐glutamic acids, followed by removal of the ester groups by dilute alkali, afforded N‐([3a,4,5,6,7,7a‐13C6]indol‐3‐ylacetyl)‐L‐aspartic and N‐([3a,4,5,6,7,7a‐13C6]indol‐3‐ylacetyl)‐L‐glutamic acids, labeled forms of compounds involved in the regulation of plant growth and development. The corresponding conjugates of (R,S)‐2,3‐dihydro‐2‐oxoindole‐3‐acetic acid, which are likewise of physiological significance, were labeled with 15N in the amino acid moieties and were synthesized via the N‐hydroxysuccinimide ester. © 1997 John Wiley & Sons, Ltd.

Phenylacetic acid metabolism in land plants: novel pathways and metabolites

“…IAA-glc and [ 13 C 6 ]IAA-glc were synthesized according to Kai et al (2007a , b ) with minor modifications. Selected l -amino acid (Val, Leu, Phe, Trp, Asp, Glu) conjugates with PAA, including isotopically labelled standards [ 13 C 6 ]PAA-Asp and [ 13 C 6 ]PAA-Glu, were prepared according to Illić et al (1997) . PAA-glc was synthesized adopting reaction conditions from Takeuchi et al (2020) .…”

Phenylacetic acid metabolism in land plants: novel pathways and metabolites

“…IAA-glc and [ 13 C 6 ]IAA-glc were synthesized according to Kai et al (2007a , b ) with minor modifications. Selected l -amino acid (Val, Leu, Phe, Trp, Asp, Glu) conjugates with PAA, including isotopically labelled standards [ 13 C 6 ]PAA-Asp and [ 13 C 6 ]PAA-Glu, were prepared according to Illić et al (1997) . PAA-glc was synthesized adopting reaction conditions from Takeuchi et al (2020) .…”

Acyl substrate preferences of an IAA-amido synthetase account for variations in grape (Vitis vinifera L.) berry ripening caused by different auxinic compounds indicating the importance of auxin conjugation in plant development

“…IAA–amino acid conjugates were synthesized according to Ilić et al (1997) using either unlabelled IAA or [indole-D 5 ]-labelled IAA as the starting substrate. The same protocol was used for the synthesis of NAA–Asp and BTOA–Asp, but since no labelled forms of NAA and BTOA were commercially available [1,4- 13 C 2 ]-L-aspartic acid was used to obtain labelled conjugates of these compounds.…”

Metabolism of exogenous auxin by Arabidopsis thaliana: Identification of the conjugate N‐(indol‐3‐ylacetyl)‐glutamine and initiation of a mutant screen

“…Indol‐3‐ylacetyl moiety: Δ 10.86 (s, 1 H, H‐1), 7.21 (1 H d, J 1,2 = 1.9 Hz, H‐2), 7.57 (1 H, d, J 4,5 =7.8 Hz, H‐4), 6.97 (1 H, t, J 5,6 =7.3 Hz, H‐5), 7.07 (1 H, t, H‐6), 7.34 (1 H, d, J 6,7 = 8.1 Hz, H‐7), 3.58 (2 H, s, CH 2 ) ppm; l ‐glutamine moiety: Δ 8.29 (1 H, d, J vic ‐1 = 7.9 Hz, NH), 7.31, 6.81 (2×1 H, 2 s, NH 2 ), 4.20 (1 H, dt, CH‐2), 1.98 (1 H, dtd, J vic ‐1 = 5.3 Hz, J vic ‐2 =7.6 Hz, J gem =13.9 Hz, CH I ‐3), 1.80 (1 H, dtd, J vic ‐1 = 6.7 Hz, J vic ‐2 = 8.7 Hz, CH II ‐3), 2.15 (2 H, t, CH 2 ‐4) ppm. Interpretation of the NMR spectra was done in comparison with spectra previously obtained for IAA‐Glu ( Ilic et al 1997).…”

Phenylacetic acid metabolism in land plants: novel pathways and metabolites

“…IAA-glc and [ 13 C 6 ]IAA-glc were synthesized according to Kai et al (2007a , b ) with minor modifications. Selected l -amino acid (Val, Leu, Phe, Trp, Asp, Glu) conjugates with PAA, including isotopically labelled standards [ 13 C 6 ]PAA-Asp and [ 13 C 6 ]PAA-Glu, were prepared according to Illić et al (1997) . PAA-glc was synthesized adopting reaction conditions from Takeuchi et al (2020) .…”

Acyl substrate preferences of an IAA-amido synthetase account for variations in grape (Vitis vinifera L.) berry ripening caused by different auxinic compounds indicating the importance of auxin conjugation in plant development

“…IAA–amino acid conjugates were synthesized according to Ilić et al (1997) using either unlabelled IAA or [indole-D 5 ]-labelled IAA as the starting substrate. The same protocol was used for the synthesis of NAA–Asp and BTOA–Asp, but since no labelled forms of NAA and BTOA were commercially available [1,4- 13 C 2 ]-L-aspartic acid was used to obtain labelled conjugates of these compounds.…”

Metabolism of exogenous auxin by Arabidopsis thaliana: Identification of the conjugate N‐(indol‐3‐ylacetyl)‐glutamine and initiation of a mutant screen

“…Indol‐3‐ylacetyl moiety: Δ 10.86 (s, 1 H, H‐1), 7.21 (1 H d, J 1,2 = 1.9 Hz, H‐2), 7.57 (1 H, d, J 4,5 =7.8 Hz, H‐4), 6.97 (1 H, t, J 5,6 =7.3 Hz, H‐5), 7.07 (1 H, t, H‐6), 7.34 (1 H, d, J 6,7 = 8.1 Hz, H‐7), 3.58 (2 H, s, CH 2 ) ppm; l ‐glutamine moiety: Δ 8.29 (1 H, d, J vic ‐1 = 7.9 Hz, NH), 7.31, 6.81 (2×1 H, 2 s, NH 2 ), 4.20 (1 H, dt, CH‐2), 1.98 (1 H, dtd, J vic ‐1 = 5.3 Hz, J vic ‐2 =7.6 Hz, J gem =13.9 Hz, CH I ‐3), 1.80 (1 H, dtd, J vic ‐1 = 6.7 Hz, J vic ‐2 = 8.7 Hz, CH II ‐3), 2.15 (2 H, t, CH 2 ‐4) ppm. Interpretation of the NMR spectra was done in comparison with spectra previously obtained for IAA‐Glu ( Ilic et al 1997).…”

Phenylacetic acid metabolism in land plants: novel pathways and metabolites

“…IAA-glc and [ 13 C 6 ]IAA-glc were synthesized according to Kai et al (2007a , b ) with minor modifications. Selected l -amino acid (Val, Leu, Phe, Trp, Asp, Glu) conjugates with PAA, including isotopically labelled standards [ 13 C 6 ]PAA-Asp and [ 13 C 6 ]PAA-Glu, were prepared according to Illić et al (1997) . PAA-glc was synthesized adopting reaction conditions from Takeuchi et al (2020) .…”

Acyl substrate preferences of an IAA-amido synthetase account for variations in grape (Vitis vinifera L.) berry ripening caused by different auxinic compounds indicating the importance of auxin conjugation in plant development

“…IAA–amino acid conjugates were synthesized according to Ilić et al (1997) using either unlabelled IAA or [indole-D 5 ]-labelled IAA as the starting substrate. The same protocol was used for the synthesis of NAA–Asp and BTOA–Asp, but since no labelled forms of NAA and BTOA were commercially available [1,4- 13 C 2 ]-L-aspartic acid was used to obtain labelled conjugates of these compounds.…”

Metabolism of exogenous auxin by Arabidopsis thaliana: Identification of the conjugate N‐(indol‐3‐ylacetyl)‐glutamine and initiation of a mutant screen

“…Indol‐3‐ylacetyl moiety: Δ 10.86 (s, 1 H, H‐1), 7.21 (1 H d, J 1,2 = 1.9 Hz, H‐2), 7.57 (1 H, d, J 4,5 =7.8 Hz, H‐4), 6.97 (1 H, t, J 5,6 =7.3 Hz, H‐5), 7.07 (1 H, t, H‐6), 7.34 (1 H, d, J 6,7 = 8.1 Hz, H‐7), 3.58 (2 H, s, CH 2 ) ppm; l ‐glutamine moiety: Δ 8.29 (1 H, d, J vic ‐1 = 7.9 Hz, NH), 7.31, 6.81 (2×1 H, 2 s, NH 2 ), 4.20 (1 H, dt, CH‐2), 1.98 (1 H, dtd, J vic ‐1 = 5.3 Hz, J vic ‐2 =7.6 Hz, J gem =13.9 Hz, CH I ‐3), 1.80 (1 H, dtd, J vic ‐1 = 6.7 Hz, J vic ‐2 = 8.7 Hz, CH II ‐3), 2.15 (2 H, t, CH 2 ‐4) ppm. Interpretation of the NMR spectra was done in comparison with spectra previously obtained for IAA‐Glu ( Ilic et al 1997).…”