title: Functional prothrombinase complex assembly on isolated monocytes and lymphocytes.
abstract: Isolated peripheral blood monocytes and lymphocytes interact with Factor Va and Factor Xa to form a functional catalytic complex which proteolytically activates prothrombin to thrombin. The kinetics of prothrombin activation were monitored continuously using the fluorescent, reversible thrombin inhibitor, dansylarginine N-(3-ethyl-1,5-pentanediyl)amide, which displays enhanced fluorescence upon binding to thrombin. Incubation of monocytes or lymphocytes with prothrombin, the cofactor (Factor Va), and the enzyme (Factor Xa) in the presence of Ca2+ generated thrombin at rates/cell exceeding those previously obtained with either bovine or human platelets. The rate of thrombin generation by monocytes exceeded that of lymphocytes and increased as monocytes adhered to a surface. Monocyte prothrombinase activity appears to be mediated through interactions, whereby Factor Va forms a receptor for Factor Xa at the monocyte surface. Monocytes possess approximately 16,100 Factor Va binding sites with a dissociation constant (Kd) of 4 X 10(-11) M. In addition, isolated, well washed monocytes and lymphocytes, respectively, contain approximately 61,400 +/- 9,900 and 24,500 +/- 4,800 molecules of Factor V/cell as determined by radioimmunoassay. Bioassay data of mononuclear cell preparations paralleled the radioimmunoassay data. The Factor V associated with washed mononuclear cells appears to be intracellular and not membrane-associated. The release of Factor V, and perhaps other sequestered coagulation factors, by these immunoreactive cells at an inflammatory site, coupled with the ability of these cells to effect thrombin generation may explain the relationship between extravascular fibrin deposition and mononuclear cell accumulation in the pathogenesis of inflammatory lesions.
mesh_terms: D001792:Blood Platelets / Q000201:enzymology; D005165:Factor V / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D015943:Factor Va; D005170:Factor X / Q000096:biosynthesis* / Q000302:isolation & purification; D015951:Factor Xa; D006801:Humans; D007700:Kinetics; D008214:Lymphocytes / Q000201:enzymology*; D008970:Molecular Weight; D009000:Monocytes / Q000201:enzymology*; D011516:Prothrombin / Q000378:metabolism
chemical_list: C022475:prothrombinase complex; D015943:Factor Va; D005165:Factor V; D011516:Prothrombin; D005170:Factor X; D015951:Factor Xa