Purification and partial amino acid sequence of the glutamate 1-semialdehyde aminotransferase of barley and synechococcus.
Glutamate-1-semialdehyde aminotransferase (E.C. 5.4.3.8) was purified from barley and the cyanobacteria Synechococcus PCC 6301. The purification procedure involved serial affinity chromatography and preparative polyacrylamide gel electrophoresis under non-denaturing conditions. The aminotransferase of these two organisms showed different mobilities in non-denaturing gels. In SDS-PAGE the enzyme from both organisms migrated as a single protein with an apparent molecular weight of 46.000 Da. An antibody against the barley enzyme cross-reacted with the cyanobacterial aminotransferase. This antibody also recognized a 17 kDa peptide cleaved from the barley protein with cyanogen bromide. Amino acid sequences of the NH2-termini revealed significant homology between the eucaryotic and cyanobacterial enzyme.
Grimm
B
B
Department of Physiology, Carlsberg Laboratory, Copenhagen Valby.
Bull
A
A
Welinder
K G
KG
Gough
S P
SP
Kannangara
C G
CG
D000595:Amino Acid Sequence; D015153:Blotting, Western; D002736:Chloroplasts / Q000201:enzymology; D000458:Cyanobacteria / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D001467:Hordeum / Q000201:enzymology; D019751:Intramolecular Transferases; D007525:Isoelectric Focusing; D007700:Kinetics; D008969:Molecular Sequence Data; D010944:Plants / Q000201:enzymology*; D012689:Sequence Homology, Nucleic Acid; D000637:Transaminases / Q000302:isolation & purification* / Q000378:metabolism
D000637:Transaminases; D019751:Intramolecular Transferases; C061049:glutamate-1-semialdehyde 2,1-aminomutase