Glutamine synthetase isoenzymes of Phaseolus vulgaris L.: subunit composition in developing root nodules and plumules

Abstract

In the legume Phaseolus vulgaris L., glutamine synthetase (GS) (EC.6.3.1.2.) occurs as three cytosolic polypeptides, α, β and γ, and a plastidic polypeptide, δ. This paper describes the subunit composition of active octameric GS isoenzymes from root nodules and plumules using ionexchange high-performance liquid chromatography followed by two-dimensional denaturing gel electrophoresis and Western immunodetection. Root nodules contained four separable GS activities, three of which were composed mainly of cytosolic γ, γ/β and β GS polypeptides, whereas the fourth activity, consisted of plastidic δ GS polypeptides. The increase in GS activity during nodulation was due largely to the appearance of γ-containing isoenzymes, and to a lesser extent on the δ isoenzyme, whereas the β-isoenzyme activity remained approximately constant throughout. Plumule GS from imbibed seeds was found to be composed of separate α and β isoenzymes, but 2 d after germination, plumule GS consisted of a mixture of α, α/β and β isoenzymes. The results from both nodules and plumules indicate that different cytosolic GS polypeptides in P. vulgaris are able to assemble into both homo-octameric and heterooctameric isoenzymes. Moreover, the changes in the patterns of isoenzymes observed during nodule development and plumule growth are interpreted to be caused both by temporal changes in the denovo synthesis of the polypeptides and also by their spatial separation in different cell types.