Rotational dynamics of spin-labelled surfactant-associated proteins SP-B and SP-C in dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylglycerol bilayers.
Pulmonary surfactant proteins SP-B and SP-C have been isolated from porcine lungs and selectively labelled with 2,2,6, 6-tetramethylpiperidine-N-oxyl (TEMPO)-isothiocyanate at their N-terminal amine ends, to analyse the mobility of both proteins on the nanosecond time scale using electron spin resonance (ESR) spectroscopy. Reconstitution of the labelled forms of these proteins in bilayers of dipalmitoylphosphatidylcholine (DPPC) or dipalmitoylphosphatidylglycerol (DPPG) results in much broader and anisotropic ESR spectra, indicating a large restriction in rotational mobility of the protein-attached probe when inserted in membranes. Distinctive differences were found between the ESR spectra of the two polypeptides, that were consistent with intrinsic differences in mode of interaction of SP-B and SP-C with phospholipid bilayers. The mobility of the protein spin probes was sensitive to temperature on the time scale of conventional spin-label ESR. Both proteins, TEMPO-SP-B and TEMPO-SP-C, showed considerable increases in mobility at temperatures above the pretransition of pure DPPC. Finally, the mobility of the spin probes attached to both SP-B and SP-C was more restricted in DPPG than in DPPC bilayers, demonstrating that electrostatic interactions of the positively charged residues at the protein surface influence the rotational dynamics of the proteins in anionic lipid bilayers. Although some residual segmental mobility of the thiourea-linked probes cannot be discounted, the results clearly reflect preferential differences in overall protein dynamics in gel and fluid phases of the two phospholipids that could be important for the biophysical properties of surfactant bilayers and monolayers.
Cruz
A
A
Departamento Bioquímica, Facultad Biología, Universidad Complutense, 28040 Madrid, Spain.
Marsh
D
D
Pérez-Gil
J
J
D015060:1,2-Dipalmitoylphosphatidylcholine / Q000737:chemistry*; D004578:Electron Spin Resonance Spectroscopy; D008051:Lipid Bilayers / Q000737:chemistry*; D010715:Phosphatidylglycerols / Q000737:chemistry*; D011510:Proteolipids / Q000737:chemistry*; D011663:Pulmonary Surfactants / Q000737:chemistry*; D013113:Spin Labels; D013696:Temperature
D008051:Lipid Bilayers; D010715:Phosphatidylglycerols; D011510:Proteolipids; D011663:Pulmonary Surfactants; D013113:Spin Labels; D015060:1,2-Dipalmitoylphosphatidylcholine; C030345:1,2-dipalmitoylphosphatidylglycerol