2 アズキ黄化芽生え中に含まれるジベレリン結合タンパク質の精製

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We have reported the characteristics and partial purification of a gibberellin-binding protein (GBP) in Azuki bean seedlings. This GBP showed specific binding to active Gas. Here we report further purification of the GBP where a reversed phase perfusion chromatography was crucial. After extracion of buffer-soluble proteins, precipitation of the protein by salting-out, and by polyethyleneimine treatment, the crude protein fraction was purified by three successive ion exchange column chromatographies as reported beore, and a fraction showing GA-binding activity was obtained. The active fraction thus obtained was subjected to a reversed phase active fraction showing a larger retention time was collected, concentrated, and subjected to a gel-filtration HPLC of SMART system. Checking the elution pattern of GA-binding activitiy, we denatured the major and the adjacent side fractions with TFA, and subjected it to a reversed phase HPLC of SMART system. After careful examination of the changes of the UV'-absorption profile and GA-binding activity in each fraction, we could detect one peak whose intensity of UV-absorption (280nm) and GA-binding activity changed together between before and after TFA treatment. We are collecting the peak for sequencing as a good candidate of GBP.
植物化学調節学会の論文
1999-10-01