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Articles

A Comparison of Two Sucrose Synthetase Isozymes from Normal and shrunken-1 Maize ¹

USDA-ARS, University of Florida, Gainesville, Florida 32611, Department of Plant Pathology, University of Florida, Gainesville, Florida 32611, Department of Agronomy, University of Florida, Gainesville, Florida 32611

The maize sucrose synthetase isozyme (SS2) present in sh1 endosperm, sh1 seedlings, and in suspension culture cells was purified to homogeneity from each of these tissues by sequential ammonium sulfate fractionation, diethylaminoethyl-cellulose chromatography, gel filtration chromatography, and affinity elution with UTP from a carboxymethyl-cellulose column. Cyanogen bromide digests were used to demonstrate that the SS2 isozymes in these different tissues are structurally identical and are therefore the product of the same gene. The sucrose synthetase produced by the Sh1 gene (SS1) was purified by modification of the SS2 procedure and was used in comparative analyses of the two isozymes. Ouchterlony assays demonstrated that SS1 and SS2 have partial antigenic identity. The two isozymes have similar enzyme kinetics in the sucrose cleavage reaction but differ in their relative activities with ADP and TDP. The amino acid compositions of SS1 and SS2 are similar, and proteolytic digests revealed that they share limited structural homologies.

¹ Cooperative investigation, Agricultural Research Service, United States Department of Agriculture and Institute of Food and Agricultural Sciences, University of Florida. Florida Agricultural Experiment Station Journal Series No. 6323. Supported in part by the United States Department of Agriculture, Competitive Research Grants Office, Grant No. 82-CRCR-1-1064.

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