Factor Va-membrane interaction is mediated by two regions located on the light chain of the cofactor.
Factor Va was incubated with 1-azidopyrene, a fluorescent lipophilic probe, in the presence of phospholipid vesicles composed of various proportions of phosphatidylcholine (PC) and phosphatidylserine (PS). The majority of the label was associated with the light chain of factor Va. The light chain was found to be labeled in the presence of phospholipid vesicles containing either 100% PC or 100% PS. After cleavage by factor Xa and incubation with PC/PS vesicles composed of 75% PC and 25% PS, label was found both on the M(r) = 30,000 fragment, derived from the NH2-terminal portion of the bovine factor Va light chain (residues 1537-1752), and on the M(r) = 46,000/48,000 carboxyl-terminal fragment of the factor Va light chain (residues 1753-2183). The M(r) = 46,000/48,000 fragment incorporated 1-azidopyrene independent of the phospholipid composition, while label incorporation into the M(r) = 30,000 fragment required phospholipid vesicles containing PC. No labeling of the M(r) = 30,000 fragment was observed with phospholipid vesicles composed of 100% PS. The label incorporation into the two portions of the molecule was found to be independent of the ionic strength in the presence of phospholipid vesicles containing 75% PC and 25% PS. In contrast, the labeling of the M(r) = 46,000/48,000 fragment with phospholipid vesicles composed of 100% PS was ionic strength dependent.(ABSTRACT TRUNCATED AT 250 WORDS)
Kalafatis
M
M
Department of Biochemistry, University of Vermont, College of Medicine, Burlington 05405-0068.
Rand
M D
MD
Mann
K G
KG
D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D001665:Binding Sites; D002417:Cattle; D004492:Edetic Acid / Q000494:pharmacology; D015943:Factor Va / Q000378:metabolism*; D005456:Fluorescent Dyes; D008081:Liposomes / Q000378:metabolism*; D008970:Molecular Weight; D009994:Osmolar Concentration; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D010713:Phosphatidylcholines / Q000032:analysis / Q000378:metabolism; D010718:Phosphatidylserines / Q000032:analysis / Q000378:metabolism; D011721:Pyrenes / Q000378:metabolism
D017304:Annexin A5; D005456:Fluorescent Dyes; D008081:Liposomes; D010446:Peptide Fragments; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D011721:Pyrenes; C028591:azidopyrene; D015943:Factor Va; D004492:Edetic Acid