Placental anticoagulant proteins: isolation and comparative characterization four members of the lipocortin family.
Previously we isolated and characterized a placental anticoagulant protein (PAP or PAP-I), which is a Ca2+-dependent phospholipid binding protein [Funakoshi et al. (1987) Biochemistry 26, 5572] and a member of the lipocortin family [Funakoshi et al. (1987) Biochemistry 26, 8087]. In this study, three additional anticoagulant proteins (PAP-II, PAP-III, and PAP-IV) were simultaneously isolated from human placental homogenates prepared in the presence of 5 mM ethylenediaminetetraacetic acid. The isoelectric points of PAP-I, PAP-II, PAP-III, and PAP-IV were 4.8, 6.1, 5.9, and 8.1, respectively, and their apparent molecular weights were 32,000, 33,000, 34,000, and 34,500, respectively. Amino acid sequences of cyanogen bromide fragments of these proteins showed that PAP-III was a previously unrecognized member of the lipocortin family, while PAP-II was probably the human homologue of porcine protein II and PAP-IV was a derivative of lipocortin II truncated near the amino terminus. Comparative studies showed that all four proteins inhibited blood clotting and phospholipase A2 activity with potencies consistent with their measured relative affinities for anionic phospholipid vesicles. However, PAP-IV bound to phospholipid vesicles approximately 160-fold more weakly than PAP-I, while PAP-II and PAP-III bound only 2-fold and 3-fold more weakly. These results increase to six the number of lipocortin-like proteins known to exist in human placenta. The observed differences in phospholipid binding may indicate functional differences among the members of the lipocortin family despite their considerable structural similarities.
Tait
J F
JF
Department of Biochemistry, University of Washington, Seattle 98195.
Sakata
M
M
McMullen
B A
BA
Miao
C H
CH
Funakoshi
T
T
Hendrickson
L E
LE
Fujikawa
K
K
D000595:Amino Acid Sequence; D017302:Annexins; D000925:Anticoagulants / Q000302:isolation & purification*; D003488:Cyanogen Bromide; D005260:Female; D006023:Glycoproteins / Q000302:isolation & purification*; D006801:Humans; D008081:Liposomes; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D010713:Phosphatidylcholines; D010740:Phospholipases / Q000037:antagonists & inhibitors*; D010741:Phospholipases A / Q000037:antagonists & inhibitors*; D054467:Phospholipases A2; D010920:Placenta / Q000378:metabolism*; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000302:isolation & purification*
D017302:Annexins; D000925:Anticoagulants; D006023:Glycoproteins; D008081:Liposomes; D010446:Peptide Fragments; D010713:Phosphatidylcholines; D011257:Pregnancy Proteins; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; D003488:Cyanogen Bromide