3-Oxoacyl-(acyl-carrier protein) reductase from avocado (Persea americana) fruit mesocarp.
The NADPH-linked 3-oxoacyl-(acyl-carrier protein) (ACP) reductase (EC 1.1.1.100), also known as 'beta-ketoacyl-ACP reductase', has been purified from the mesocarp of mature avocado pears (Persea americana). The enzyme is inactivated by low ionic strength and low temperature. On SDS/PAGE under reducing conditions, purified 3-oxoacyl-ACP reductase migrated as a single polypeptide giving a molecular mass of 28 kDa. Gel-filtration chromatography gave an apparent native molecular mass of 130 kDa, suggesting that the enzyme is tetrameric. The enzyme is inactivated by dilution, but some protection is afforded by the presence of NADPH. Kinetic constants have been determined using synthetic analogues as well as the natural ACP substrate. It exhibits a broad pH optimum around neutrality. Phenylglyoxal inactivates the enzyme, and partial protection is given by 1 mM-NADPH. Antibodies have been raised against the protein, which were used to localize it using immunogold electron microscopy. It is localized in plastids. N-Terminal amino-acid-sequence analysis was performed on the enzyme, and it shows close structural similarity with cytochrome f. Internal amino-acid-sequence data, derived from tryptic peptides, shows similarity with the putative gene products encoded by the nodG gene from the nitrogen-fixing bacterium Rhizobium meliloti and the gra III act III genes from Streptomyces spp.
Sheldon
P S
PS
Department of Biochemistry, University of Birmingham, U.K.
Kekwick
R G
RG
Sidebottom
C
C
Smith
C G
CG
Slabas
A R
AR
D064431:3-Oxoacyl-(Acyl-Carrier-Protein) Reductase; D000096:Acetone / Q000031:analogs & derivatives / Q000494:pharmacology; D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004795:Enzyme Stability; D005638:Fruit / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera / Q000276:immunology; D007460:Iodoacetamide / Q000494:pharmacology; D007700:Kinetics; D016253:Microscopy, Immunoelectron / Q000379:methods; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010658:Phenylglyoxal / Q000494:pharmacology; D011817:Rabbits; D013347:Subcellular Fractions / Q000201:enzymology
D007106:Immune Sera; D000096:Acetone; C047048:1,3-dibromo-2-propanone; D000429:Alcohol Oxidoreductases; D064431:3-Oxoacyl-(Acyl-Carrier-Protein) Reductase; D010658:Phenylglyoxal; D007460:Iodoacetamide