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231224s2016 xx |||||o 00| ||eng c |
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|a 10.1016/j.plaphy.2016.05.031
|2 doi
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|a pubmed25n0869.xml
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|a (DE-627)NLM260829641
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|a (NLM)27235647
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|a (PII)S0981-9428(16)30201-7
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|a DE-627
|b ger
|c DE-627
|e rakwb
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| 041 |
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|a eng
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| 100 |
1 |
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|a Ostrowski, Maciej
|e verfasserin
|4 aut
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| 245 |
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|a Cloning and biochemical characterization of indole-3-acetic acid-amino acid synthetase PsGH3 from pea
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|c 2016
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|a Text
|b txt
|2 rdacontent
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|a ƒaComputermedien
|b c
|2 rdamedia
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|a ƒa Online-Ressource
|b cr
|2 rdacarrier
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|a Date Completed 27.03.2017
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|a Date Revised 09.01.2024
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|a published: Print-Electronic
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|a Citation Status MEDLINE
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|a Copyright © 2016 Elsevier Masson SAS. All rights reserved.
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|a Phytohormone conjugation is one of the mechanisms that maintains a proper hormonal homeostasis and that is necessary for the realization of physiological responses. Gretchen Hagen 3 (GH3) acyl acid amido synthetases convert indole-3-acetic acid (IAA) to IAA-amino acid conjugates by ATP-dependent reactions. IAA-aspartate (IAA-Asp) exists as a predominant amide conjugate of auxin in pea tissues and acts as an intermediate during IAA catabolism. Here we report a novel recombinant indole-3-acetic acid-amido synthetase in Pisum sativum. In silico analysis shows that amino acid sequence of PsGH3 has the highest homology to Medicago truncatula GH3.3. The recombinant His-tag-PsGH3 fusion protein has been obtained in E. coli cells and is a soluble monomeric polypeptide with molecular mass of 69.18 kDa. The PsGH3 was purified using Ni(2+)-affinity chromatography and native PAGE. Kinetic analysis indicates that the enzyme strongly prefers IAA and L-aspartate as substrates for conjugation revealing Km(ATP) = 0.49 mM, Km(L-Asp) = 2.2 mM, and Km(IAA) = 0.28 mM. Diadenosine pentaphosphate (Ap5A) competes with ATP for catalytic site and diminishes the PsGH3 affinity toward ATP approximately 1.11-fold indicating Ki = 8.5 μM. L-Tryptophan acts as an inhibitor of IAA-amido synthesizing activity by competition with L-aspartate. Inorganic pyrophosphatase (PPase) hydrolyzing pyrophosphate to two phosphate ions, potentiates IAA-Asp synthetase activity of PsGH3. Our results demonstrate that PsGH3 is a novel enzyme that is involved in auxin metabolism in pea seeds
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|a Journal Article
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|a Auxin conjugate
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|a Gretchen Hagen 3
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|a Indole-3-acetic acid
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|a Indole-3-acetyl-aspartate
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|a Pisum sativum
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|a Dinucleoside Phosphates
|2 NLM
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| 650 |
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|a Enzyme Inhibitors
|2 NLM
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| 650 |
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|a Indoleacetic Acids
|2 NLM
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| 650 |
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|a Plant Proteins
|2 NLM
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| 650 |
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|a Recombinant Proteins
|2 NLM
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| 650 |
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7 |
|a Aspartic Acid
|2 NLM
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| 650 |
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|a 30KYC7MIAI
|2 NLM
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| 650 |
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|a P(1),P(5)-di(adenosine-5'-)pentaphosphate
|2 NLM
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| 650 |
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|a 50304-44-4
|2 NLM
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| 650 |
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|a indoleacetic acid
|2 NLM
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| 650 |
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|a 6U1S09C61L
|2 NLM
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| 650 |
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|a Tryptophan
|2 NLM
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| 650 |
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|a 8DUH1N11BX
|2 NLM
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| 650 |
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|a Adenosine Triphosphate
|2 NLM
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| 650 |
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7 |
|a 8L70Q75FXE
|2 NLM
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| 650 |
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|a Ligases
|2 NLM
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| 650 |
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|a EC 6.-
|2 NLM
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| 700 |
1 |
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|a Mierek-Adamska, Agnieszka
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Porowińska, Dorota
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Goc, Anna
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Jakubowska, Anna
|e verfasserin
|4 aut
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| 773 |
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|i Enthalten in
|t Plant physiology and biochemistry : PPB
|d 1991
|g 107(2016) vom: 15. Okt., Seite 9-20
|w (DE-627)NLM098178261
|x 1873-2690
|7 nnns
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| 773 |
1 |
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|g volume:107
|g year:2016
|g day:15
|g month:10
|g pages:9-20
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| 856 |
4 |
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|u http://dx.doi.org/10.1016/j.plaphy.2016.05.031
|3 Volltext
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|a AR
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|d 107
|j 2016
|b 15
|c 10
|h 9-20
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