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Glutamine synthetases of green and etiolated leaves ofSinapis alba

Evidence of the identity of the respective enzyme proteins

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Abstract

Studies on the glutamine synthetases (GS, EC 6.3.1.2) of green (GS2) and etiolated leaves (GSet) ofSinapis alba L. (cv. Steinacher) revealed striking similarities between the respective enzyme proteins. The enzymes showed corresponding chromatographic properties, both on dimethylaminoethyl-Sephacel and on hydroxylapatite columns. The purified GS proteins were also identical with regard to the molecular weight of their subunits. Isoelectrofocusing of pure GSet yielded two distinct polypeptide bands in the pH 5.6 region of the gels. This pattern corresponded to the two strong bands of GS2. Two charge variants of GS polypeptides could be detected by Western-blot analysis of the soluble protein of green leaves using antibodies against mustard GS2. In immunoprecipitation experiments, the holoenzymes of GS2 and GSet were recognized with identical affinities by this antiserum. We conclude that strong similarities exist between the proteins of the GS enzymes in green and etiolated leaves of mustard. Most probably only one GS form, namely the plastidic enzyme, can be found in the epigeal organs ofSinapis. The polypeptides of the GS2 subunits showed no differences in the hydrophobicity of the polypeptide chains. Neither glucosyl nor mannosyl residues could be detected.

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Abbreviations

DEAE:

diethylaminoethyl

GS, GS1, GS2, GSet :

glutamine synthetase, cytosolic GS, plastidic GS, GS from etiolated leaves

IgG:

immunoglobulin G

2-ME:

2-mercaptoethanol

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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Authors and Affiliations

  1. Institut für Allgemeine Botanik der Johannes Gutenberg-Universität, Saarstrasse 21, D-6500, Mainz, Germany

    M. Höpfner, G. Ochs & A. Wild

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  1. M. Höpfner
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  2. G. Ochs
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  3. A. Wild
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Dedicated to Professor Dr. H. Mohr on the occasion of his 60th birthday

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Höpfner, M., Ochs, G. & Wild, A. Glutamine synthetases of green and etiolated leaves ofSinapis alba . Planta 181, 155–161 (1990). https://doi.org/10.1007/BF02411532

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  • DOI: https://doi.org/10.1007/BF02411532

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