Distribution of phospholipid transfer protein in human plasma: presence of two forms of phospholipid transfer protein, one catalytically active and the other inactive.
Plasma phospholipid transfer protein (PLTP) plays an important role in the maintenance of plasma high-density lipoprotein (HDL) content and remodeling of HDL in the circulation. In the present study we have used different fractionation methods to investigate the distribution of PLTP in human plasma. A novel enzyme-linked immunosorbent assay developed during the study allowed for simultaneous assessment of both PLTP mass and activity in the fractions obtained. Size-exclusion chromatography and plasma fractionation by nondenaturing polyacrylamide gel electrophoresis (PAGE) yielded similar results demonstrating that PLTP associates in native plasma with two distinct particle populations, while ultracentrifugation with high salt leads to detachment of PLTP from lipoprotein particles and loss of a majority of its phospholipid transfer activity. Interestingly, analysis of the size-exclusion chromatography fractions demonstrated that PLTP exists in the circulation as an active population that elutes in the position of HDL corresponding to an average molecular mass of 160+/-40 kDa and an inactive form with an average mass of 520+/-120 kDa. The inactive fraction containing approximately 70% of the total PLTP protein eluted between HDL and low density lipoprotein (LDL). Thus, the two PLTP pools are associated with different types of lipoprotein particles, suggesting that the PLTP activity in circulation is modulated by the plasma lipoprotein profile and lipid composition.
Oka
T
T
Research Department, Research and Development Center, BML, Kawagoe, Saitama 350-1101, Japan.
Kujiraoka
T
T
Ito
M
M
Egashira
T
T
Takahashi
S
S
Nanjee
M N
MN
Miller
N E
NE
Metso
J
J
Olkkonen
V M
VM
Ehnholm
C
C
Jauhiainen
M
M
Hattori
H
H
D000911:Antibodies, Monoclonal; D016632:Apolipoprotein A-I / Q000737:chemistry / Q000378:metabolism; D016633:Apolipoprotein A-II / Q000737:chemistry / Q000378:metabolism; D015153:Blotting, Western; D002352:Carrier Proteins / Q000097:blood* / Q000737:chemistry / Q000276:immunology; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007527:Isoenzymes; D008075:Lipoproteins, HDL / Q000737:chemistry / Q000378:metabolism; D008565:Membrane Proteins / Q000097:blood* / Q000737:chemistry / Q000276:immunology; D008970:Molecular Weight; D010316:Particle Size; D049294:Phospholipid Transfer Proteins; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D014461:Ultracentrifugation
D000911:Antibodies, Monoclonal; D016632:Apolipoprotein A-I; D016633:Apolipoprotein A-II; D002352:Carrier Proteins; D007527:Isoenzymes; D008075:Lipoproteins, HDL; D008565:Membrane Proteins; D049294:Phospholipid Transfer Proteins; D011994:Recombinant Proteins