Chloroplastic glutamine synthetase is activated by direct binding of aluminium.
Acidification of soils may release water soluble, toxic aluminium species from clay minerals. Al interferes with a wide range of physical and cellular processes. Glutamine synthetase (GS, EC 6.3.1.2) is the key enzyme of primary N assimilation, as well as ammonia reassimilation and detoxification. Plant GS requires two magnesium ions per subunit for activity, which makes GS a potential target of metal stress. The objective of this investigation was to prove that Al from an organic metal complex is able to activate GS, and Al becomes bound to the polypeptide structure of the GS molecule. Aluminium(III)-nitrilotriacetic acid complex (Al(III)NTA) activated the GS prepared from wheat (Triticum aestivum L.) leaves, as Al(3+) did in vivo, but could not functionally substitute magnesium ions, which were also necessary for the activity in the in vitro GS assay. GS2 was isolated by non-denaturing polyacrylamide gel electrophoresis, and the Al and Mg content of the enzyme was determined by inductively coupled plasma atomic emission spectroscopy. The GS octamer remained intact and contained Mg(2+) bound to its specific sites after the electrophoretic separation. Al was detected in the Al(III)NTA-treated sample bound to the structure of the enzyme protein, potentially occupying one of the specific metal-binding sites of the subunits. Our results indicate that the activatory effect of the Al(III)NTA complex is because of specific binding of aluminium to the polypeptide chain of GS2, however presence of magnesium at least on one of the metal-binding sites is essential to the active state of the enzyme.
Pécsváradi
Attila
A
Department of Plant Biology, University of Szeged, Szeged, Hungary. pecsvaradi@bio.u-szeged.hu
Nagy
Zoltán
Z
Varga
Anikó
A
Vashegyi
Agnes
A
Labádi
Imre
I
Galbács
Gábor
G
Zsoldos
Ferenc
F
D000535:Aluminum / Q000378:metabolism*; D002736:Chloroplasts / Q000201:enzymology*; D020536:Enzyme Activators / Q000378:metabolism*; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism; D009571:Nitrilotriacetic Acid / Q000378:metabolism; D018515:Plant Leaves / Q000201:enzymology; D010940:Plant Proteins / Q000378:metabolism; D014908:Triticum / Q000201:enzymology*
D020536:Enzyme Activators; D010940:Plant Proteins; D000535:Aluminum; C083818:glutamine synthetase 2; D005974:Glutamate-Ammonia Ligase; D008274:Magnesium; D009571:Nitrilotriacetic Acid