| Abstract: | Soluble acid invertase from wheat coleoptiles was purified toelectrophoretic homogeneity. A comparison of molecular weightby SDS-PAGE and gel filtration suggested that the enzyme wasa monomer of Mr50 000. The enzyme was a glycoprotein and, afterchemical deglycosylation, possessed a Mrof 48000. A polyclonalantiserum was raised against the deglycosylated protein. Thiscross-reacted specifically with acid invertase. A putative precursorof invertase synthesized in a cell-free translation system wasdetected by SDS-PAGE and fluorography of the immunoprecipitatedpolypeptides. The distribution of acid invertase in wheat seedlingshoots was investigated both by visualizing invertase activityafter starch gel electrophoresis and by immunoblotting. Bothtechniques identified two forms of invertase in extracts ofthe primary leaf and only one form in extracts of coleoptiles.The low pH optimum and the glycoprotein nature of wheat coleoptileinvertase are consistent with a vacuolar location. Fructoseinhibited its activity, suggesting that enzyme activity couldbe modulated by end-product inhibition. Key words: Acid invertase, purification, antiserum, glycoprotein, Triticum aestivum, wheat, coleoptiles |
