Plant Physiol. Drug Metab Dispos
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Plant Physiology 76:871-878 (1984)
© 1984 American Society of Plant Biologists

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Articles

Subcellular Localizations of Two Dolichos biflorus Lectins 1

Marilynn E. Etzler, Susan MacMillan, Steven Scates2, Donna M. Gibson3, Douglas W. James, Jr.4, Douglas Cole and Susan Thayer

Department of Biochemistry and Biophysics, University of California, Davis, California 95616

The subcellular localizations of the Dolichos biflorus seed lectin and the structurally related lectin (cross-reactive material [CRM]) from the stems and leaves of this plant were determined by immunofluorescence, immunocytochemistry, and cell fractionation procedures. Subcellular fractionation of the cotyledons using a nonaqueous procedure to minimize disruption of the protein bodies showed that the majority of the seed lectin was associated with the protein body fraction and some lectin was also present in the starch granules. Immunofluorescence and immunocytochemistry at the light microscopic level showed that the seed lectin was mainly localized at the peripheries of these organelles. Lectin was also found in the cytoplasm of the cells, although the amount appeared to be dependent upon the degree of protein body disruption.

Immunofluorescence and immunocytochemistry studies of the stem and leaf lectin (CRM) indicated that a significant portion of this lectin may be associated with the cell walls, although lectin was also seen in the cytoplasm of plasmolyzed cells. Extraction and cell fractionation studies showed that a large portion of the CRM is readily solubilized and most of the remainder is pelleted at 1000g. The CRM can be extracted from these pellets by treatment with cellulase and pectinase; other reagents such as NaCl, detergents, and EDTA could also release significant amounts of CRM. These studies suggest that the CRM is noncovalently bound to the cell walls. A comparison of the distribution of exogenously supplied [125I]CRM with the endogenous CRM during extraction and cell fractionation indicates that soluble CRM is not adsorbed to the 1000g pellet during fractionation.

The different subcellular distributions of these two structurally related lectins suggest that different tissues of the same plant may utilize lectins for different functions.


2 Present address: Washington University School of Medicine, St. Louis, MO 63110.

3 Present address: Frito-Lay Inc., Basic Research, 900 North Loop 12, Irving, TX 75061.

4 Present address: Advanced Genetics Science, Inc., 6701 San Pablo Ave., Oakland, CA 94608.

1 Supported by United States Public Health Services Grant GM 21882, United States Department of Agriculture Grant 79-59-2063-1-1-242, and National Science Foundation Grant PCM8215758.







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Copyright © 1984 by the American Society of Plant Biologists