Partial purification and photoaffinity labelling of sunflower acyl-CoA:lysophosphatidylcholine acyltransferase.
Previous attempts to purify acyl-CoA:1-acyl-lysophosphatidylcholine acyltransferase (EC 2.3.1.23) have been frustrated by difficulties in solubilizing the enzyme without inactivation. Microsomal preparations, from the developing cotyledons of sunflower, in high concentrations of urea retain activity. Gel-filtration liquid chromatography followed by trypsin treatment (minus urea) resulted in the removal of many contaminating proteins without loss of enzyme activity. SDS/PAGE showed the presence of two major peptides with apparent molecular masses of 52 and 59 kDa. These polypeptides cross-reacted with the radiolabelled photoreactive substrate 1-azido-oleoyl-sn-lysophosphatidyl-[N-methyl-(3)H]choline.
Fraser
T
T
School of Biological Sciences, University of Bristol, Woodlands Road, Bristol BS8 1UG, UK. t.c.m.fraser@bristol.ac.uk
Stobart
K
K
D008243:1-Acylglycerophosphocholine O-Acyltransferase / Q000302:isolation & purification* / Q000378:metabolism*; D000345:Affinity Labels; D000818:Animals; D002850:Chromatography, Gel; D006368:Helianthus / Q000201:enzymology*; D007425:Intracellular Membranes / Q000201:enzymology; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology*; D008862:Microsomes, Liver / Q000201:enzymology; D010782:Photolysis; D051381:Rats; D013379:Substrate Specificity; D014466:Ultraviolet Rays
D000345:Affinity Labels; D008243:1-Acylglycerophosphocholine O-Acyltransferase