Secretable human platelet-derived factor V originates from the plasma pool

被引:86
作者
Camire, RM
Pollak, ES
Kaushansky, K
Tracy, PB
机构
[1] Univ Vermont, Coll Med, Dept Biochem, Burlington, VT 05405 USA
[2] Childrens Hosp, Div Hematol, Philadelphia, PA 19104 USA
[3] Univ Washington, Sch Med, Div Hematol, Seattle, WA 98195 USA
关键词
D O I
10.1182/blood.V92.9.3035.421k54_3035_3041
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Factor Va (FVa), derived from plasma or released from stimulated platelets, is the essential protein cofactor of the prothrombinase complex. Plasma-derived factor V (FV) is synthesized by the liver, whereas the source of the platelet-derived cofactor has not been unambiguously identified. Megakaryocytes, platelet precursors, are known to synthesize platelet proteins and to endocytose proteins from plasma (ie, fibrinogen) and then package these proteins into alpha-granules. io determine which mechanism accounts for RI presence in platelets, two patients heterozygous for FVLeiden who underwent allogeneic transplantation from homozygous RI wild-type donors (bone marrow [BM] or liver) were studied. Patient JMW, whose skin biopsy specimen showed heterozygous FVLeiden, received a BM transplant from a wild-type homozygous RV donor as analyzed from posttransplant peripheral blood cells. Patient RN,whose native liver is heterozygous for FVLeiden, received a homozygous wild-type FV liver. Because each individual has two distinct genetic pools of factor V in liver and megakaryocytes, it was possible to determine whether secretable platelet-derived RI was normal or contained the FVLeiden mutation. Platelet-derived FVa released from thrombin-activated platelets from a normal individual, an individual heterozygous for the FVLeiden mutation, and the two patients was incubated with phospholipid vesicles and activated protein C (APC). Western blotting analyses using a monoclonal antibody that allows distinction between platelet-derived FVa and FVa(Leiden) subsequent to APC-catalyzed cleavage were then performed. Based on the accumulation of proteolytic fragments derived from APC-induced cleavage, analyses of platelet-derived FVa from JMW demonstrated both normal FVa and FVa(Leiden) consistent with a plasma-derived origin of the secretable platelet-derived FVa. Western blotting analyses of the APC-cleaved platelet-derived FVa from FW showed a wild-type phenotype, despite the presence of a FVLeiden allele in her megakaryocyte genome, also consistent with a plasma origin of her secretable platelet-derived FVa. Platelets do not appear to endocytose the plasma cofactor, because a 35-hour incubation of platelet-rich plasma with I-125-factor V showed no specific association/uptake of the radiolabeled ligand with the platelet pellet. Collectively, these results show for the first time that the majority of secretable platelet-derived factor V is endocytosed by megakaryocytes from plasma and is not exclusively synthesized by these cells, as previously believed. (C) 1998 by The American Society of Hematology.
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页码:3035 / 3041
页数:7
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