Conversion of glycerate to serine in intact spinach leaf peroxisomes.
Intact spinach (Spinacia oleracea L.) leaf peroxisomes converted glycerate to serine in the presence of NAD and alanine. The reaction proceeded optimally at pH9. Addition of oxaloacetate or alpha-ketoglutarate plus aspartate enhanced the conversion about three-fold. Alteration of the concentration of one of the reaction components, consisting of 2 mM glycerate, 0.2 mM NAD, 0.5 mM oxaloacetate, and 2 mM alanine, revealed half-saturation constants of 0.45 mM for glycerate, 0.06 mM for NAD, 0.02 mM for oxaloacetate, and 0.33 mM for alanine. The conversion proceeded with the formation of hydroxypyruvate followed by serine; hydroxypyruvate did not accumulate to a high amount in the presence or absence of alanine. The amino group donor could be alanine (half-saturation constant, 0.33 mM), glycine (0.45 mM), or asparagine (0.67 mM); the three amino acids produced roughly similar Vmax values. The results indicate that, in the conversion of glycerate to serine, the transamination is catalyzed by a hydroxypyruvate aminotransferase with characteristics unknown among all other studied leaf peroxisomal aminotransferases. The peroxisomal membrane is sparsely permeable to NAD/NADH, and the participation of the peroxisomal malate dehydrogenase in an electron shuttle system across the membrane in the regeneration of NAD/NADH is suggested.
D000409:Alanine / Q000378:metabolism; D000429:Alcohol Oxidoreductases / Q000378:metabolism; D001711:Biotransformation; D055598:Chemical Phenomena; D002621:Chemistry; D005988:Glyceric Acids / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D050537:Hydroxypyruvate Reductase; D007700:Kinetics; D008830:Microbodies / Q000378:metabolism*; D009243:NAD / Q000378:metabolism; D010071:Oxaloacetates / Q000378:metabolism; D010944:Plants / Q000378:metabolism*; D012694:Serine / Q000096:biosynthesis*
D005988:Glyceric Acids; D010071:Oxaloacetates; D009243:NAD; D012694:Serine; D000429:Alcohol Oxidoreductases; D050537:Hydroxypyruvate Reductase; D000409:Alanine