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The specificities of a human plasma and bovine liver phospholipid transfer protein were studied using a fluorescence assay based on the transfer of pyrenyl phospholipids. This method was used previously to determine the mechanism of spontaneous transfer of phospholipids between model lipoproteins (Massey, J.B., Gotto, A.M., Jr. and Pownall, H.J. (1982) Biochemistry 21, 3630-3636). The pyrenyl phospholipids varied in the headgroup moiety; pyrenyl phosphatidylcholines contained different fatty acyl chains in the sn-1 position. Model high-density lipoproteins (R-HDL) consisting of apolipoprotein A-I and 1-palmitoyl-2-oleoylphosphatidylcholine (POPC) were used as donor and acceptor particles. As previously shown, the bovine liver protein mediated the transfer of only phosphatidylcholine. In contrast, the human plasma protein transferred all species studied which included a phosphatidylserine, phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, phosphatidic acid, sphingomyelin, galactosylcerebroside, and a diacylglycerol. The activity of these transfer proteins was only slightly affected by changes in the acyl chain composition of the transferring lipid. Pyrenyl and radioactive ([3H]POPC) phospholipids were transferred with equal rates by the human transfer protein, suggesting that this protein has similar binding characteristics for pyrenyl and natural phospholipids. Spontaneous phospholipid transfer occurs by the aqueous diffusion of monomeric lipid where the rate is highly dependent on fatty acyl chain composition. In this study, no correlation between the rate of spontaneous transfer and protein-mediated transfer was found. The apparent Km values for R-HDL and low-density lipoprotein (LDL), when used as acceptors, were similar when based on the number of acceptor particles. The apparent Vmax for the bovine liver protein was identical for R-HDL and LDL but for the plasma protein Vmax was slightly higher for R-HDL. These results suggest that, like the bovine liver protein, the plasma protein functions as a phospholipid-binding carrier that exchanges phospholipids between membrane surfaces. The assay of lipid transfer proteins by pyrenyl-labeled lipids is faster and easier to perform than other current methods, which require separation of donor and acceptor particles, and is suitable for studies on the function and mechanism of action of lipid transfer proteins.
J B Massey, and
D Hickson-Bick, and
D P Via, and
A M Gotto, and
H J Pownall
October 1980,
Biochimica et biophysica acta,
J B Massey, and
D Hickson-Bick, and
D P Via, and
A M Gotto, and
H J Pownall
July 1985,
Biochimica et biophysica acta,
J B Massey, and
D Hickson-Bick, and
D P Via, and
A M Gotto, and
H J Pownall
March 1997,
Biochemistry,
J B Massey, and
D Hickson-Bick, and
D P Via, and
A M Gotto, and
H J Pownall
May 1986,
Biochimica et biophysica acta,
J B Massey, and
D Hickson-Bick, and
D P Via, and
A M Gotto, and
H J Pownall
October 1996,
Biochimica et biophysica acta,
J B Massey, and
D Hickson-Bick, and
D P Via, and
A M Gotto, and
H J Pownall
December 1980,
Biochimica et biophysica acta,
J B Massey, and
D Hickson-Bick, and
D P Via, and
A M Gotto, and
H J Pownall
May 1992,
Biochimica et biophysica acta,
J B Massey, and
D Hickson-Bick, and
D P Via, and
A M Gotto, and
H J Pownall
September 1999,
Biology of reproduction,
J B Massey, and
D Hickson-Bick, and
D P Via, and
A M Gotto, and
H J Pownall
January 1988,
Advances in experimental medicine and biology,
J B Massey, and
D Hickson-Bick, and
D P Via, and
A M Gotto, and
H J Pownall
August 2010,
Biochemistry,
