Identification of alpha-1 acid glycoprotein as a lysophospholipid binding protein: a complementary role to albumin in the scavenging of lysophosphatidylcholine.
Alpha-1 acid glycoprotein (AGP, orosomucoid), a major acute phase protein in plasma, displays potent cytoprotective and anti-inflammatory activities whose molecular mechanisms are largely unknown. Because AGP binds various exogenous drugs, we have searched for endogenous ligands for AGP. We found that AGP binds lysophospholipids in a manner discernible from albumin in several ways. First, mass spectrometric analyses showed that AGP isolated from plasma and serum contained lysophosphatidylcholine (LPC) enriched in mono and polysaturated acyl chains, whereas albumin contained mostly saturated LPC. Second, AGP bound LPC in a 1:1 molar ratio and with a higher affinity than free fatty acids, whereas albumin bound LPC in a 3:1 ratio but with a lower affinity than that of free fatty acids. Consequently, free fatty acids displaced LPC more avidly from albumin than from AGP. Competitive ligand displacement indicated the highest affinity for AGP to LPC20:4, 18:3, 18:1, and 16:0 (150-180 nM), lysophosphatidylserine (Kd 190 nM), and platelet activating factor (PAF) (Kd 235 nM). The high affinity of AGP to LPC in equilibrium was verified by stopped-flow kinetics, which implicated slow dissociation after fast initial binding, being consistent with an induced-fit mechanism. AGP also bound pyrene-labeled phospholipids directly from vesicles and more efficiently than albumin. AGP prevented LPC-induced priming and PAF-induced activation of human granulocytes, thus indicating scavenging of the cellular effects of the lipid ligands. The results suggest that AGP complements albumin as a lysophospholipid scavenging protein, particularly in inflammatory conditions when the capacity of albumin to sequester LPC becomes impaired.
Ojala
Pauli J
PJ
Finnish Red Cross Blood Service, Kivihaantie 7, FI-00310 Helsinki, Finland. pauli.ojala@bts.redcross.fi
Hermansson
Martin
M
Tolvanen
Martti
M
Polvinen
Kirsi
K
Hirvonen
Tia
T
Impola
Ulla
U
Jauhiainen
Matti
M
Somerharju
Pentti
P
Parkkinen
Jaakko
J
D005456:Fluorescent Dyes; D008246:Lysophospholipids / Q000378:metabolism*; D013058:Mass Spectrometry; D008958:Models, Molecular; D009961:Orosomucoid / Q000737:chemistry / Q000378:metabolism*; D010767:Phosphorylcholine / Q000378:metabolism*; D011485:Protein Binding
D005456:Fluorescent Dyes; D008246:Lysophospholipids; D009961:Orosomucoid; D010767:Phosphorylcholine