The Location of Glutamine Synthetase in Leaf Cells and its Role in the Reassimilation of Ammonia Released in Photorespiration

The localization of glutamine synthetase within the cells ofbarley and pea leaves has been reinvestigated using either amechanical technique or rupturing of isolated protoplasts torelease the cellular organelles, and both differential and densitygradient centrifugation to separate them. In no case could wefind evidence for any significant association between glutaminesynthetase and the mitochondria; our results suggest that theenzyme is present in the chloroplast and the cytoplasm of bothspecies. Experiments with isolated mitochondria from spinachalso failed to provide any suggestion that these organellesmight contain glutamine synthetase. Thus there is no evidenceto support the hypothesis, published by others, that mitochondriareassimilate ammonia, released from glycine oxidation, by meansof their own glutamine synthetase. Further experiments werecarried out to see if glutamate dehydrogenase present in themitochondria could reassimilate ammonia under conditions inwhich the electron transport chain to oxygen was blocked. Althoughthere was some evidence for a small amount of assimilatory glutamatedehydrogenase activity under these conditions it was dependenton adding relatively high concentrations of ammonia and wasinsufficient to sustain the rate of recycling of NAD requiredfor glycine oxidation. The results were thus considered to becompatible with the operation of the photorespiratory nitrogencycle as previously published.