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Plant Physiology 79:34-40 (1985)
© 1985 American Society of Plant Biologists

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Articles

Starch Branching Enzymes from Maize 1

Immunological Characterization using Polyclonal and Monoclonal Antibodies

Bijay K. Singh and Jack Preiss2

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

Spleen cells from mice immunized with starch branching enzymes were fused with cells from the mouse myeloma Sp2/0-AG14 cell line to form hybridomas. Those hybridomas producing antibodies against the branching enzyme were screened by the enzyme-linked immunosorbent assay using purified branching enzyme as the antigen. Three monoclonal cell lines (1A1D7, 1A1C3 and 4D2A9D8) were found to produce antibodies which showed positive enzyme-linked immunosorbent assay reactions with maize branching enzyme I in addition to branching enzymes IIa and IIb. Three other monoclonal cell lines (4D2D10, 4D2F9, and 2A6C12) were also selected which were found to produce antibodies showing positive enzyme-linked immunosorbent assay reactions with branching enzymes IIa and IIb only.

Amino acid composition and peptide maps obtained after trypsin or chymotrypsin digestion show that there is no difference between branching enzyme IIa and IIb but they are significantly different from branching enzyme I which, along with immunological data, suggests that only two forms of starch branching enzyme may be present in maize kernels.

Immunological cross-reaction was also found between the starch branching enzyme from maize kernels and the glycogen branching enzyme from Escherichia coli using polyclonal antibodies against starch branching enzyme I or IIa and IIb or E. coli glycogen branching enzyme, suggesting some immunological similarities between maize starch branching enzymes and E. coli glycogen branching enzyme.

2 Present address: Department of Biochemistry, Michigan State University, East Lansing, MI 48824.

1 Supported in part by National Science Foundation grant PCM 82-05705.




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S. Rahman, A. Regina, Z. Li, Y. Mukai, M. Yamamoto, B. Kosar-Hashemi, S. Abrahams, and M. K. Morell
Comparison of Starch-Branching Enzyme Genes Reveals Evolutionary Relationships Among Isoforms. Characterization of a Gene for Starch-Branching Enzyme IIa from the Wheat D Genome Donor Aegilops tauschii
Plant Physiology, March 1, 2001; 125(3): 1314 - 1324.
[Abstract] [Full Text]


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Copyright © 1985 by the American Society of Plant Biologists