Abstract
An auxin binding sive, with characteristics different from the previously described auxin binding sites I and II in maize coleoptiles, is reported in homogenates of zucchini (Cucurbita pepo L. cv. Black Beauty) hypocotyls. Evidence from differential centrifugation and sucrose and metrizamide density gradients indicates that the site is localized on the plasma membrane. The site has a KD of 1–2×10−6 M for indole acetic acid and has a pH optimum of 5.0. Binding specificity measured with several auxins, weak auxins, and anti-auxins generally parallels the activities of the same compounds as inhibitors of auxin transport. 1-N-naphthylphthalamic acid and 2,3,5-triiodobenzoic acid (2,3,5-TIBA), both auxin transport inhibitors in vivo, increase specific auxin binding to this site. 3,4,5-TIBA, which can partially reverse 2,3,5-TIBA's transport inhibition when the two substances are added together in vivo, partially reverses 2,3,5-TIBA's increase in specific auxin binding to the plasma membrane site when added with 2,3,5-TIBA in vitro. Preliminary investigations indicate that a similar plasma membrane site exists in maize (Zea mays L.) coleoptiles. It is suggested that different conformations of this site may function during active auxin transport.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- IAA:
-
indole-3-acetic acid
- NPA:
-
1-N-naphthylphthalamie acid
- 2,3,5-TIBA:
-
2,3,5-triiodobenzoic acid
- 3,4,5-TIBA:
-
3,4,5-triiodobenzoic acid
- 1-NAA:
-
1-naphthaleneacetic acid
- 2-NAA:
-
2-naphthaleneacetic acid
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- DTE:
-
dithioerythritol
- MOPS:
-
N-morpholino-3-propansulfonic acid
- CCO:
-
cytochrome c oxidase
- CCR:
-
NADH: cytochrome c reductase
- glu I:
-
glucan synthetase I
- ER:
-
endoplasmic reticulum
References
Appelmans, F.R., Wattiaux, R., De Duve, C.: Tissue fractionation studies. 5. The association of acid phosphatase with a special class of cytoplasmic granules in rat liver. Biochem. J. 59, 438–445 (1955)
Batt, S., Venis, M.A.: Separation and localization of two classes of auxin binding sites in corn coleoptile membranes. Planta 130, 15–21 (1976)
Dohrmann, U., Hertel, R., Kowalik, H.: Properties of auxin binding in different subcellular fractions from maize coleoptiles. Planta 140, 97–106 (1978)
Döllstadt, R., Hirschberg, K., Winkler, E., Hübner, G.: Bindung von Indolylessigsäure und Phenoxyessigsäure an Fraktionen aus Epikotylen und Wurzeln von Pisum sativum L. Planta 130, 105–111 (1976)
Goldsmith, M.H.M.: The polar transport of auxin. Ann. Rev. Plant Phys. 28, 439–478 (1977)
Goldsmith, M.H.M., Breault, D.: Evidence for the existence of binding sites in the polar transport of indoleacetic acid. Plant Physiol. 53, S 43 (1974)
Gross, J., Marmé, D.: ATP-dependent Ca2+ uptake into plant membrane vesicles. Proc. Nat. Acad. Sci. USA 75, 1232–1236 (1978)
Hertel, R., Evans, M.L., Leopold, A.C., Sell, H.M.: The specificity of the auxin transport system. Planta 85, 238–249 (1969)
Hill, T.L.: Analysis of a model for active transport. Proc. Natl. Acad. Sci. USA 65, 409–416 (1970)
Ihl, M.: Indole-acetic acid binding proteins in soybean cotyledon. Planta 131, 223–228 (1976)
Kasamo, K., Yamaki, T.: In vitro binding of IAA to plasma membrane-rich fractions containing Mg++-activated ATPase from mung, bean hypocotyls. Plant Cell Physiol. 17, 149–164 (1976)
Lord, J.M., Kagawa, T., Moore, T.S., Beevers, H.: Endoplasmic reticulum as the site of lecithin formation in castor bean endosperm. J. Cell Biol. 57, 659–667 (1973)
Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J.: Protein measurement with the Folin, phenol reagent. J. Biol. Chem. 193, 265–275 (1951)
Oostrom, H., Van Loopik-Detmers, M.A., Libbenga, K.R.: A high-affinity receptor for indele-acetic acid in cultured tobacco pith explants. FEBS Letters 59, 194–197 (1975)
Ray, P.M.: Regulation of β-glucan synthetase activity by auxin in pea stem tissue. Plant Physiol. 51, 601–614 (1973)
Ray, P.M.: Auxin binding, sites of maize coleoptiles are localized on membranes of the endoplasmic reticulum. Plant Physiol. 59, 594–599 (1977)
Ray, P.M., Dohrmann, U., Hertel, R.: Characterization of naphthaleneacetic acid binding to receptor sites on cellular membranes of maize coleoptile tissue. Plant Physiol. 59, 359–364 (1977a)
Ray, P.M., Dohrmann, U., Hertel, R.: Specificity of auxin-binding sites on maize coleoptile membranes as possible receptor sites for auxin action. Plant Physiol. 60, 585–591 (1977b)
Rickwood, D., Birnie, G.D.: Metrizamide: a new density-gradient medium. FEBS Letters. 50, 102–110 (1975)
Rubery, P.H., Sheldrake, A.R.: Carrier-mediated auxin transport. Planta 118, 101–121 (1974)
Rubery, P.H.: The specificity of carrier-mediated auxin transport by suspensioncultured crown gall cells. Planta 135, 275–283 (1977)
Semadeni, E.G.: Enzymatische Charakterisierung der Lysosomenäquivalente (Sphärosomen) von Maiskeimlingen Planta 72, 91–118 (1967)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Jacobs, M., Hertel, R. Auxin binding to subcellular fractions from Cucurbita hypocotyls: In vitro evidence for an auxin transport carrier. Planta 142, 1–10 (1978). https://doi.org/10.1007/BF00385113
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00385113