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J. Biochem, 1988, Vol. 104, No. 5 742-754
© 1988 Japanese Biochemical Society

research-article

Characterization of a Glucose Polymer from PC12 Cells and Neuronal Cells of Rat Embryo1

Maija-Liisa Rasilo2 and Tatsuya Yamagata3

Laboratory of Glycoconjugate Research, Mitsubishi Kasei Institute of Life Sciences Machida, Tokyo 194

3 To whom correspondence should be addressed

A large-sized glucose polymer was isolated by pronase digestion from line PC12 pheochromocytoma cells metabolically labeled with [1-3H]galactose The polymer was included on a column of concanavalin A-Sepharose and could be eluted with 10 mM methyl-{alpha}-mannoside. Its slight retention in a column of Bio-Gel A-5m suggested that its molecular weight was in the several millions. Glucose was the component monosaccharide and there were two minor lipophilic components present. The polymer was digested with {alpha}-amylase into a series of oligosaccharides and was cleaved by glucoamylase into glucose residues. The disaccharide obtained by digestion with {alpha}-amylase was identified as maltose in several HPLC systems and by NMR spectroscopy. NMR measurement revealed the trisaccharide to be maltotriose. Susceptibility of the polymer molecule to {alpha}-amylase, and the digestion products obtained, indicated a resemblance to glycogen. An analysis for saccharide compositions before and after reduction of the polymer suggested the presence of an aglycon part. Contrary to expectations based on the presence of this moiety, the polymer displayed good solubility in neutral organic solvents. Two-thirds of the glucose polymer was also soluble in 10% TCA. A similar glucose polymer was isolated from neuronal cells of rat embryos metabolically labeled with [1-3H]galactose. Mouse neuroblastoma cells did not synthesize the polymer.

1 This work was supported in part by Grants from Emil Aaltonen Foundation, The Finnish Academy of Sciences and The Finnish Academy.

2 Visiting guest researcher from Department of Biochemisiry, University of Helsinki. Present address: Department of General Chemistry, Faculty of Agriculture and Forestry, University of Helsinki, Viikki, 00710 Helsinki, Finland.


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