Lung calcium-dependent phospholipid-binding proteins: structure and function.
Distinct peptide maps of two rabbit lung Ca2(+)-dependent phospholipid-binding proteins (PLBPs), 36,000 and 33,000, were generated by cyanogen bromide (CNBr) cleavage, trypsin or Staphylococcus aureus V8 proteinase digestion. The amino acid sequence of a CNBr-cleaved peptide of the 36,000 PLBP was aligned to the amino terminus of human lipocortin I with more than 77% identity, but had no identity with the known amino terminal sequence of other known annexins. Partial amino acid sequence of a 33,000 PLBP peptide demonstrated a close (56%) relationship to endonexin II, human placental anticoagulant protein, and porcine intestine protein II, but shared only 32% identity with lipocortin I, 30% with lipocortin II. Antiserum generated against purified 36,000 PLBP reacted strongly with the 33,000 PLBP, but did not react with any other rabbit lung cytosolic proteins. Both PLBPs inhibited the phospholipase A2 reaction when dioleoyl phosphatidylcholine and phosphatidylglycerol vesicles or monolayers were used as substrates. In the vesicle assay, the phospholipase A2 reaction was inhibited at lower substrate phospholipid concentrations but not at nearly saturating substrate concentrations. In the monolayer assay, the phospholipid-binding proteins did not inhibit phospholipase A2 at a low phospholipid surface concentration of 3.8.10(-3) molecules/A2, but they did at higher surface concentrations between 1.1 x 10(-2) and 3.8 x 10(-2) molecules/A2. The inhibition of phospholipase A2 by rabbit lung phospholipid-binding proteins is most likely due to the prevention of penetration by phospholipase A2 into the interface, a requirement for the enzyme to act on the substrate.
Tsao
F H
FH
Department of Pediatrics, University of Wisconsin, Madison 53715.
Hull
W M
WM
Strickland
M S
MS
Whitsett
J A
JA
Foo
T K
TK
Zografi
G
G
DeLuca
P M
PM
D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003488:Cyanogen Bromide; D015180:Electrophoresis, Gel, Two-Dimensional; D005227:Fatty Acids / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D008168:Lung / Q000378:metabolism*; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010741:Phospholipases A / Q000378:metabolism; D054467:Phospholipases A2; D010743:Phospholipids / Q000378:metabolism*; D011817:Rabbits; D016415:Sequence Alignment; D013329:Structure-Activity Relationship; D013552:Swine
D002352:Carrier Proteins; D005227:Fatty Acids; D010743:Phospholipids; D010741:Phospholipases A; D054467:Phospholipases A2; D003488:Cyanogen Bromide; D002118:Calcium