Separation of mesophyll and bundle sheath cells (MC and BSC) from the leaves of green amaranth (Amaranthus retroflexus L.) showed that glycolate oxidase (GO, EC 22.214.171.124) is located predominately in BSC (on the average, 84.5% of the total activity). Three peaks of GO activity were detected following the elution from a DEAE-fractogel column. The first peak corresponded to the isoform located in BSC, the second peak had dual location, and the third one was associated with MC fraction. Elaborated flow sheet of GO purification from the amaranth leaves produced highly purified (by 63.5 times) isoforms from MC and BSC with specific activity of 0.54 EU/mg protein. It was also shown that GO from MC has greater affinity for glycolate, with the K M values for GO from BSC and MC being 58 and 20 µM, respectively. Intermediates of the Krebs cycle were shown to affect the GO activity from MC and BSC: succinate suppressed and isocitrate activated GO.
Russian Journal of Plant Physiology – Springer Journals
Published: Aug 9, 2005
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