A complex containing both trypsin inhibitor and dehydroascorbate reductase activities isolated from mitochondria of etiolated mung bean (Vigna radiata L. (Wilczek) cv. Tainan no. 5) seedlings.
A complex containing trypsin inhibitor (TI) activity was extracted with 0.1 M TRIS buffer (pH 7.9) from trypsin-treated mitochondria of etiolated mung bean seedlings, and further purified with a Superdex 200 FPLC column. This partially purified complex with an M(r) about 820 kDa exhibited additional dehydroascorbate (DHA) reductase activity with specific activities of 0.21, 1.53 and 1.54 mumol ascorbate formed min-1 mg-1 protein at pH 6.0, 6.5 and 7.0, respectively, when glutathione was added. Much lower DHA reductase activity (0.013 and 0.026 mumol ascorbate formed min-1 mg-1 protein at pH 6.5 and 7.0, respectively) was found when glutathione was omitted. The isolated complex gave positive results when it was tested by TI activity staining after SDS-PAGE, and could be recognized by a polyclonal antibody which was raised against 38 kDa sweet potato Kunitz-type TI, one of the root storage proteins of sweet potato. The possible physiological functions of this complex with both TI and DHA reductase activities were discussed.
Hou
W C
WC
Institute of Botany, Academia Sinica, Nankang, Taipei, Taiwan, Republic of China.
Wang
Y T
YT
Lin
Y H
YH
Hsiao
L J
LJ
Chen
T E
TE
Wang
C W
CW
Dai
H
H
D002845:Chromatography; D003683:Dehydroascorbic Acid / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D007887:Fabaceae / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D008928:Mitochondria / Q000737:chemistry* / Q000201:enzymology; D010088:Oxidoreductases / Q000302:isolation & purification*; D010940:Plant Proteins / Q000302:isolation & purification*; D010946:Plants, Medicinal; D014361:Trypsin Inhibitors; D000516:alpha-Amylases / Q000037:antagonists & inhibitors
D010940:Plant Proteins; D014361:Trypsin Inhibitors; D010088:Oxidoreductases; C020666:glutathione dehydrogenase (ascorbate); D000516:alpha-Amylases; D003683:Dehydroascorbic Acid