SYNTHESIS OF PHOSPHOLIPID-BASED DETERGENTS AND THEIR EFFECTS ON THE CA2+-ATPASE OF SARCOPLASMIC-RETICULUM

Phospholipid molecules containing a cholate or hemisuccinylcholate moiety at the 2-position were synthesized as possible detergents for solubilization of membrane proteins. 1-Myristoleoyl-2-cholyl-sn-glycero-3-phosphatidylcholine ((C14:1,cholyl)PC) was found to solubilize sarcoplasmic reticulum vesicles at concentrations above its cmc of ca. 4 mu g/ml. Effects of (C14:1,cholyl)PC on the activity of the Ca2+-ATPase of sarcoplasmic reticulum were complex, as for other detergents. High concentrations (0.2 mg/ml) of (C14:1,cholyl)PC were able to displace phospholipids from the lipid/protein interface of the ATPase. Although under these conditions the activity of the ATPase was low, the ATPase was not denatured since activity could be regained by displacement of (C14:1,cholyl)PC with the detergent C(12)E(8). 1-oleoyl-2-cholyl-sn-glycero-3-phosphatidylcholine ((C18:1,cholyl)PC) was found to have a very low water solubility, but this could be increased by the introduction of a hemisuccinyl group to give 1-oleoyl-2-(3 alpha-hemisuccinyl)cholyl-sn-glycero-3-phosphatidylcholine ((C18:1,cholylCOOH)PC). This was able to solubilize and delipidate the Ca2+-ATPase; the ATPase was stable in (C18:1,cholylCOOH)PC for long periods of time.