Thermotropic and dynamic characterization of interactions of acylated alpha-bungarotoxin with phospholipid bilayer membranes.

The interactions of palmitoyl-alpha-bungarotoxin (PBGT) with dipalmitoylphosphatidylcholine (DPPC) bilayers have been studied by using high-sensitivity differential scanning calorimetry together with steady-state and time-resolved phosphorescence and fluorescence spectroscopy. The incorporation of PBGT into large single lamellar vesicles causes a decrease in the phospholipid phase transition temperature (Tm), a broadening of the heat capacity function, and a decrease in the enthalpy change associated with the phospholipid gel to liquid-crystalline transition. Analysis of the dependence of this decreased enthalpy change on the protein/lipid molar ratio indicates that each PBGT molecule exhibits a localized effect upon the bilayer, preventing approximately six lipid molecules from participating in the lipid phase transition. Additional calorimetric experiments indicate that binding to acetylcholine receptor enriched membranes causes a small increase in the Tm of the PBGT/DPPC vesicles. Steady-state fluorescence depolarization measurements employing 1,6-diphenyl-1,3,5-hexatriene (DPH) indicate that the association of PBGT with the phospholipid bilayer decreases the apparent order of the bulk lipid below Tm while increasing the order above Tm. These results have been further supported by rotational mobility measurements of erythrosin-labeled PBGT associated with giant (about 2-micron) unilamellar vesicles composed of dielaidoylphosphatidylcholine or dioleoylphosphatidylcholine using the time-dependent decay of delayed fluorescence/phosphorescence emission anisotropy. Rotational correlation times in the submillisecond time scale (about 30 microseconds) indicate that the protein is highly mobile in the fluid phase and that below Tm the rotational mobility is only slightly restricted.(ABSTRACT TRUNCATED AT 250 WORDS)

[1]  D. Chapman,et al.  Monitoring membrane protein rotational diffusion using time-averaged phosphorescence. , 1983, Biochimica et biophysica acta.

[2]  T. Markello,et al.  Calorimetric and fluorescence characterization of interactions between cytochrome b5 and phosphatidylcholine bilayers. , 1983, Biochemistry.

[3]  R. Macdonald,et al.  Differential effects of alkali metal chlorides on formation of giant liposomes by freezing and thawing and dialysis. , 1983, Biochemistry.

[4]  Jonathan A. Cooper,et al.  The transforming proteins of Rous sarcoma virus, Harvey sarcoma virus and Abelson virus contain tightly bound lipid , 1982, Cell.

[5]  A. Magee,et al.  Fatty acid acylation of eucaryotic cell membrane proteins , 1982, Biochimica et Biophysica Acta (BBA) - Reviews on Biomembranes.

[6]  T. E. Thompson,et al.  Fusion of dipalmitoylphosphatidylcholine vesicles at 4 degrees C. , 1982, Biochemistry.

[7]  R. Cherry,et al.  Lateral and rotational diffusion of bacteriorhodopsin in lipid bilayers: experimental test of the Saffman-Delbrück equations. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[8]  C. Chang,et al.  Rotational mobility of an erythrocyte membrane integral protein band 3 in dimyristoylphosphatidylcholine reconstituted vesicles and effect of binding of cytoskeletal peripheral proteins. , 1982, Biochemistry.

[9]  B. Babbitt,et al.  A model system for studies of specific membrane interactions. , 1982, Biochemistry.

[10]  B. Hughes,et al.  Extraction of membrane microviscosity from translational and rotational diffusion coefficients. , 1982, Biophysical journal.

[11]  E. W. Heinzelmann,et al.  H-2Kk and vesicular stomatitis virus G proteins are not extensively associated in reconstituted membranes recognized by T cells. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[12]  F. Barrantes,et al.  Rotational molecular dynamics of the membrane-bound acetylcholine receptor revealed by phosphorescence spectroscopy. , 1981, European journal of biochemistry.

[13]  K. Kinosita,et al.  The effect of cytochrome oxidase on lipid chain dynamics. A nanosecond fluorescence depolarization study. , 1981, Biochimica et biophysica acta.

[14]  R. Austin,et al.  ROTATIONAL DIFFUSION OF BIOLOGICAL MACROMOLECULES BY TIME‐RESOLVED DELAYED LUMINESCENCE (PHOSPHORESCENCE, FLUORESCENCE) ANISOTROPY , 1981, Annals of the New York Academy of Sciences.

[15]  H. Petty,et al.  Lateral distribution and diffusion of the C3b receptor of complement, HLA antigens, and lipid probes in peripheral blood leukocytes. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[16]  A. Kusumi,et al.  Protein-lipid interaction in rhodopsin recombinant membranes as studied by protein rotational mobility and lipid alkyl chain flexibility measurements. , 1980, Journal of Biochemistry (Tokyo).

[17]  C. Gahmberg,et al.  Rotational diffusion of band 3 proteins in membranes from En(a-) and neuraminidase-treated normal human erythrocytes. , 1980, Biochimica et biophysica acta.

[18]  R. Cherry,et al.  Rotational and lateral diffusion of membrane proteins. , 1979, Biochimica et biophysica acta.

[19]  P. Garland,et al.  Phosphorescence depolarization and the measurement of rotational motion of proteins in membranes , 1979, FEBS letters.

[20]  J. W. Parce,et al.  Antibodies bound to lipid haptens in model membranes diffuse as rapidly as the lipids themselves. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[21]  J. Gomez-Fernandez,et al.  Protein‐lipid interactions , 2022 .

[22]  F. Goñi,et al.  Intrinsic protein—lipid interactions , 1979, FEBS letters.

[23]  T. E. Thompson,et al.  Thermotropic behavior of monoglucocerebroside--dipalmitoylphosphatidylcholine multilamellar liposomes. , 1979, Biochemistry.

[24]  P. Hartig,et al.  Fast cation flux from Torpedo californica membrane preparations: implications for a functional role for acetylcholine receptor dimers. , 1978, Biochemical and biophysical research communications.

[25]  T. E. Thompson,et al.  Studies on the anomalous thermotropic behavior of aqueous dispersions of dipalmitoylphosphatidylcholine-cholesterol mixtures. , 1978, Biochemistry.

[26]  G. Shipley,et al.  Protein-lipid interactions: recombinants of the proteolipid apoprotein of myelin with dimyristoyllecithin. , 1977, Biochemistry.

[27]  Y. Barenholz,et al.  Enveloped viruses as model membrane systems: microviscosity of vesicular stomatitis virus and host cell membranes. , 1976, Biochemistry.

[28]  P. Saffman,et al.  Brownian motion in biological membranes. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[29]  Y. Wolman,et al.  Use of esters of N-hydroxysuccinimide in the synthesis of N-acylamino acids. , 1967, Journal of lipid research.

[30]  F. Greenwood,et al.  Preparation of Iodine-131 Labelled Human Growth Hormone of High Specific Activity , 1962, Nature.

[31]  C. Y. Lee,et al.  Chromatographic separation of the venom of Bungarus multicinctus and characterization of its components. , 1972, Journal of chromatography.