Gel to liquid crystalline phase transition promotes a conformational reorganization of Ca2+, Mg2+-ATPase from sarcoplasmic reticulum in dimyristoylphosphatidylcholine reconstituted systems.
Sarcoplasmic reticulum Ca2+, Mg2+-ATPase has been reconstituted in membranes highly enriched in dimyristoylphosphatidylcholine. According to electron microscopy data these membranes form vesicles of an average diameter of 1000 +/- 200 A. These reconstituted membranes show hysteretic behavior in some physical-chemical properties, such as light scattering and fluorescence when labeled with iodoacetamidofluorescein and with N-iodoacetyl-N'-(5-sulfo-1-naphthyl) ethylenediamine. Hysteretic behavior in catalytic activity can also be inferred from the kinetic data presented in this paper, because the temperature dependence of the Ca2+, Mg2+-ATPase activity is altered by a mild thermal pretreatment of the samples. Furthermore, it was noticed that the Ca2+-dependent ATPase activity of these complexes, when assayed above the phase transition temperature (Tc) of the lipid matrix, showed a lag phase in the minute time scale range. On the basis of these findings, it is suggested that the gel-to-liquid crystalline phase transition of the lipid is able to shift the conformational equilibrium E----E* of Ca2+, Mg2+-ATPase. The fact that the -SH reactivity against 5,5'-dithio-bis-nitrobenzoic acid of these complexes is also altered by preincubation above Tc for several minutes also supports that lipid melting induces a conformational change in Ca2+, Mg2+-ATPase.
D017301:Ca(2+) Mg(2+)-ATPase / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004134:Dimyristoylphosphatidylcholine; D005614:Freeze Fracturing; D007700:Kinetics; D008027:Light; D008051:Lipid Bilayers / Q000378:metabolism; D008081:Liposomes; D008854:Microscopy, Electron; D010316:Particle Size; D011487:Protein Conformation; D012519:Sarcoplasmic Reticulum / Q000201:enzymology*; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence; D013696:Temperature
D008051:Lipid Bilayers; D008081:Liposomes; D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases; D004134:Dimyristoylphosphatidylcholine