Synthesis and use of a lysolecithin analog for the purification of UDP-glucuronosyltransferase.
Because of their high cost, lysolecithins are generally not considered useful detergents for the purification of membrane-bound enzymes. Therefore, we have synthesized a structural analog of lysolecithin with similar physical properties for which synthesis is straightforward. This analog is 1-palmitoylpropanediol-3-phosphocholine. To compare the efficacy of the two detergents for the purification of a membrane-bound enzyme, we have purified UDP-glucuronosyltransferase from pig liver microsomes using lysophosphatidylcholine or the synthetic analog. The catalytic properties of UDP-glucuronosyltransferase purified with 1-palmitoylpropanediol-3-phosphocholine or lysolecithin were identical. Sodium dodecyl sulfate-gel electrophoresis indicated that the purity of the UDP-glucuronosyltransferase preparation was the same whether lysophosphatidylcholine or its synthetic analog was used. The advantage of using 1-palmitoylpropanediol-3-phosphocholine in preference to lysophosphatidylcholine is that the former can be synthesized for about 1% the cost of the latter. In addition, the method for synthesis of 1-palmitoylpropanediol-3-phosphocholine is general in that the structural features of the polymethylene chain can be varied, allowing for the inexpensive synthesis of a series of detergents.
Dannenberg
A
A
Department of Medicine, Cornell University Medical College, New York, New York 10021.
Wong
T
T
Zakim
D
D
Eibl
H
H
D004591:Electrophoresis, Polyacrylamide Gel; D014453:Glucuronosyltransferase / Q000302:isolation & purification*; D008244:Lysophosphatidylcholines / Q000138:chemical synthesis / Q000378:metabolism*
D008244:Lysophosphatidylcholines; D014453:Glucuronosyltransferase