Abstract
The 2S albumin from the endosperm of castor seed (Ricinus communis L.) seed was reduced by thioredoxin from either wheat germ or Escherichia coli. The 2S protein is made up of a large (approx. 7 kDa) subunit that contains two intramolecular disulfides and a small (approx. 4 kDa) subunit that lacks intramolecular disulfides. The two subunits are joined by at least one intermolecular disulfide bond. Thioredoxin could be reduced either enzymically with NADPH and NADP-thioredoxin reductase or chemically with dithiothreitol. Reduced glutathione and glutaredoxin (from E. coli) were without effect. The ability of the 2S protein to undergo reduction by thioredoxin was demonstrated by a direct reduction procedure based on the fluorescent probe, monobromobimane, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and by an enzymatic procedure in which reduction is linked to activation of chloroplast NADP-malate dehydrogenase. Analyses indicated that thioredoxin actively reduced the intramolecular disulfides of the 2S large subunit, but was ineffective in reducing the intermolecular disulfide(s) that connect the large to the small subunit. These findings extend the role of thioredoxin to the reduction of a seed protein that is widely distributed in oil producing plants.
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Abbreviations
- DDT:
-
dithiothreitol
- mBBr:
-
monobromobimane
- NTR:
-
NADP-thioredoxin reductase
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
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This work was supported by a grant from the National Science Foundation.
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Shin, S., Wong, J.H., Kobrehel, K. et al. Reduction of castor-seed 2S albumin protein by thioredoxin. Planta 189, 557–560 (1993). https://doi.org/10.1007/BF00198219
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DOI: https://doi.org/10.1007/BF00198219