Infrared spectroscopic study of a "hard-to-exchange" phospholipid population in the vicinity of CaATPase.
CaATPase from rabbit sarcoplasmic reticulum (SR) has been isolated, purified, and reconstituted into unilamellar vesicles with 1,2-dipentadecanoylphosphatidylcholine (DPePC) and acyl-chain-perdeuterated 1,2-dipalmitoylphosphatidylcholine (DPPC-d62). For total lipid:protein mole ratios between 25 and 70, a constant "hard-to-exchange" phospholipid population (HEPP) of 12 +/- 4 native phospholipid molecules per protein monomer is observed, consistent with the studies of Bick et al. [(1991) Arch. Biochem. Biophs. 286, 346-352]. Thermotropic and conformational properties of the lipids in native SR and in the reconstituted systems were probed with FT-IR spectroscopy. The native SR phospholipids undergo a broad phase transition centered at about 1-2 degrees C and are thus disordered under physiological conditions. The thermotropic behavior of CH2 wagging progressions characteristic of the palmitate chains differs from that of the total lipid population and is suggestive of membrane microheterogeneity. The individual thermotropic and conformational properties of the HEPP and the exogenous lipid in reconstituted vesicles containing CaATPase and DPPC-d62 were monitored. At temperatures below the onset of the gel-liquid-crystal phase transition, the HEPP possesses significant conformational disorder, and exhibits the monotonic introduction of gauche rotamers as the temperature is raised from -55 to 27 degrees C, in contrast to the exogenous lipid, which exhibits a constant high order over the same temperature range. Nevertheless, the HEPP undergoes a residual order-disorder phase transition with similar but not identical parameters (half-width, midpoint temperature) to the gel-liquid-crystal transition of the bulk (exogenous) lipid in the reconstituted systems.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Chemistry, Newark College of Arts and Science, Rutgers University, New Jersey 07102.
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Senak
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Mendelsohn
D000818:Animals; D000252:Calcium-Transporting ATPases / Q000737:chemistry*; D005583:Fourier Analysis; D009132:Muscles / Q000201:enzymology; D010743:Phospholipids / Q000737:chemistry*; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000201:enzymology; D013055:Spectrophotometry, Infrared
D010743:Phospholipids; D000252:Calcium-Transporting ATPases