Glutamine synthetase isolated from human brain: octameric structure and homology of partial primary structure with human liver glutamine synthetase

Glutamine synthetase (GS) has been purified from the cytosolic fraction of non-frozen human brain tissue. The purified GS migrated as a main band around 44 kD on reducing SDS-PAGE. Two-dimensional electrophoresis revealed heterogeneity within subunits of GS. The masses of eight different peptides from a tryptic digest of GS as measured by high resolution MALDI-MS matched with the respective masses from an in silico tryptic fingerprint of the Swiss-Prot database entry of human liver GS, proving that at least 24% of the primary sequences of GS from brain and liver are identical. Sedimentation equilibrium profiles obtained from analytical ultracentrifugation experiments at 10°C showed that human brain GS is mainly octameric. The quaternary structure of human brain GS at 10 M (subunit concentration) was not significantly affected by cations, such as magnesium (5 and 20 mM) or manganese (0.2 and 1 mM) within the range of pH 7.1-7.8.