Photoaffinity-labeled Auxins

Leonard, Nelson J.; Greenfield, John C.; Schmitz, Ruth Y.; Skoog, Folke

doi:10.1104/pp.55.6.1057

Cited by 19 publications

(6 citation statements)

References 16 publications

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“…Several azido analogues of phytohormones have been synthesized (35,36) and their potential value in identifying plant hormone-binding proteins has recently been recognized. A major problem with the use of azido ligands in plant extracts was nonspecific background labeling (37).…”

Section: Discussionmentioning

confidence: 99%

An auxin-binding protein is localized to the plasma membrane of maize coleoptile cells: identification by photoaffinity labeling and purification of a 23-kda polypeptide.

Feldwisch

Zettl

Hesse

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

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Plasma membrane vesicles were isolated from maize (Zea mays L.) coleoptile tissue by aqueous twophase partitioning and assayed for homogeneity by the use of membrane-specific enzymatic assays. Using 5-azido- [7-3H]indole-3-acetic acid ([3H]N3IAA), we identified several IAAbinding proteins with molecular masses of 60 kDa (pm6O), 58 kDa (pm58), and 23 kDa (pm23). Using Triton X-114, we were able to selectively extract pm23 from the plasma membrane. We show that auxins and functional analogues compete with[3H]N3IAA for binding to pm23. We found that PAB130, a polyclonal antibody raised against auxin-binding protein 1 (ABP-1), recognized ABP-1 as well as pm23. This suggests that pm23 shares common epitopes with ABP-1. In addition, we identified an auxin-binding protein with a molecular mass of 24 kDa (pm24), which was detected in microsomal but not in plasma membrane vesicle preparations. Like pm23 this protein was extracted from membrane vesicles with Triton X-1 14. We designed a purification scheme allowing simultaneous purification of pm23 and pm24. Homogeneous pm23 and pm24 were obtained from coleoptile extracts after 7000-fold purification.

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Section: Discussionmentioning

confidence: 99%

An auxin-binding protein is localized to the plasma membrane of maize coleoptile cells: identification by photoaffinity labeling and purification of a 23-kda polypeptide.

Feldwisch

Zettl

Hesse

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

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“…Indole-3-acetic acid (IAA), the main auxin in higher plants, has profound effects on plant growth and development . The synthetic diazirinyl IAA derivatives ( 17a and 17b ) were subjected to oat coleoptile segment growth bioassays . The typical auxin responses, which were growth acceleration at optimum concentration and growth inhibition at higher and lower concentration than optimum concentration, were observed for the synthetic compounds.…”

Section: Resultsmentioning

confidence: 99%

Comprehensive Synthesis of Photoreactive (3-Trifluoromethyl)diazirinyl Indole Derivatives from 5- and 6- Trifluoroacetylindoles for Photoaffinity Labeling

Murai

Masuda²,

Sakihama

et al. 2012

J. Org. Chem.

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“…Since photoaffinity labelling has been a powerful tool for identification of numerous ligand-binding proteins, synthetic phytohormone analogues carrying photolabile groups had been synthesized as early as 1975 (Leonard et al, 1975). Azido-IAA derivatives were synthesized by Melhado et al (1982) and demonstrated to have auxin activity (Melhado et al, 1984;Jones et al, 1984a).…”

Section: Discussionmentioning

confidence: 99%

Identification of a 23 kDa protein from maize photoaffinity-labelled with 5-azido-[7-3H]indol-3-ylacetic acid

Zettl¹,

Campos²,

Palme³

1995

Biochemical Journal

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A 23 kDa protein (p23) was identified in microsomal extracts from maize coleoptiles by photoaffinity labelling with 5-azido-[7-3H]indol-3-ylacetic acid ([3H]N3IAA). Labelling of p23 was blocked by unlabelled IAA, N3IAA, indol-3-ylbutyric acid and indol-3-yl-lactate. In addition, labelling was efficiently decreased by tryptophan, as well as by the scavenger p-aminobenzoic acid. Labelling was, however, not affected by synthetic auxins such as 1-naphthylacetic acid or 2,4-dichlorophenoxyacetic acid. Competition data suggest that the label was probably bound via the indole ring, and hence labelling was not specific for auxins. The 23 kDa protein was solubilized from crude microsomes by extraction with Triton X-100 and purified to homogeneity by ion-exchange, size-exclusion and reversed-phase chromatography. After electroblotting, the amino acid sequences of the p23 N-terminus as well as the several tryptic peptides were obtained. Database comparisons revealed sequence identity with a maize manganese superoxide dismutase. We conclude that photoaffinity labelling of p23 was pseudo-affinity, and therefore the binding site for IAA is not specific.

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Photoaffinity-labeled Auxins

Cited by 19 publications

References 16 publications

An auxin-binding protein is localized to the plasma membrane of maize coleoptile cells: identification by photoaffinity labeling and purification of a 23-kda polypeptide.

An auxin-binding protein is localized to the plasma membrane of maize coleoptile cells: identification by photoaffinity labeling and purification of a 23-kda polypeptide.

Comprehensive Synthesis of Photoreactive (3-Trifluoromethyl)diazirinyl Indole Derivatives from 5- and 6- Trifluoroacetylindoles for Photoaffinity Labeling

Identification of a 23 kDa protein from maize photoaffinity-labelled with 5-azido-[7-3H]indol-3-ylacetic acid

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