SUBUNIT STRUCTURE OF LYSINE SENSITIVE ASPARTATE KINASE FROM SPINACH LEAVES

The lysine -sensitive isoenzyme of aspartate kinase was purified to ho mogeneity from spinach leaves and its subunit composition was studied. The purified preparation had an apparent molecular mass of 280,000 an d separated into two subunits- a large subunit with molecular mass of 53000 and smaller subunit with molecular mass of 17,000 by urea treatm ent and SDS PAGE. The enzyme molecule has subunit composition of 4 lar ge and 4 small subunits. The activity of the large subunit was stimula ted more than two fold by the addition of small subunit and the stimul ated activity was inhibited by EGTA. This inhibition could be reversed by Ca++. Further characteristics of the smaller subunit such as heat stability, behavior on ion exchange chromatography, electrophoretic mo bility on polyacrylamide gels, amino acid composition and pattern, pre sence of trimethyl lysine, its ability to activate other calmodulin st imulated enzymes and its calmodulin-like nature in RIA tests suggested that this subunit is identical to calmodulin.