PMID- 22235940
OWN - NLM
STAT- MEDLINE
DCOM- 20120830
LR  - 20230829
IS  - 1538-7836 (Electronic)
IS  - 1538-7836 (Linking)
VI  - 10
IP  - 5
DP  - 2012 May
TI  - Factor seven activating protease (FSAP): does it activate factor VII?
PG  - 859-66
LID - 10.1111/j.1538-7836.2012.04619.x [doi]
AB  - BACKGROUND: Factor seven activating protease (FSAP) was initially reported as an 
      activator of single-chain urokinase-type plasminogen activator (scuPA) and factor 
      VII (FVII). Subsequently, numerous additional substrates have been identified, 
      and multiple other biological effects have been reported. Due to the apparent 
      lack of specificity, the physiological role of FSAP has become increasingly 
      unclear. Rigorous studies have been limited by the difficulty of obtaining intact 
      FSAP from blood or recombinant sources. OBJECTIVES: Our aim was to produce intact 
      recombinant human FSAP, and to assess its role as a trigger of coagulation and 
      fibrinolysis. RESULTS: Expression of wild-type FSAP in various mammalian cells 
      invariably resulted in the accumulation of degraded FSAP due to autoactivation 
      and degradation. To overcome this problem, we constructed a variant in which 
      Arg(313) at the natural activation site was replaced by Gln, creating a cleavage 
      site for the bacterial protease thermolysin. HEK293 cells produced FSAP(R313Q) in 
      its intact form. Thermolysin-activated FSAP displayed the same reactivity toward 
      the substrate S-2288 as plasma-derived FSAP, and retained its ability to activate 
      scuPA. Polyphosphate and heparin increased V(max) by 2-3-fold, without affecting 
      K(m) (62 nm) of scuPA activation. Surprisingly, FVII activation by activated FSAP 
      proved negligible, even in the presence of calcium ions, phospholipid vesicles 
      and recombinant soluble tissue factor. On membranes of 100% cardiolipin FVII 
      cleavage did occur, but this resulted in transient activation and rapid 
      degradation. CONCLUSIONS: While FSAP indeed activates scuPA, FVII appears 
      remarkably resistant to activation. Therefore, reappraisal of the putative role 
      of FSAP in hemostasis seems appropriate.
CI  - (c) 2012 International Society on Thrombosis and Haemostasis.
FAU - Stavenuiter, F
AU  - Stavenuiter F
AD  - Department of Plasma Proteins, Sanquin Research, Amsterdam, the Netherlands.
FAU - Dienava-Verdoold, I
AU  - Dienava-Verdoold I
FAU - Boon-Spijker, M G
AU  - Boon-Spijker MG
FAU - Brinkman, H J M
AU  - Brinkman HJ
FAU - Meijer, A B
AU  - Meijer AB
FAU - Mertens, K
AU  - Mertens K
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - J Thromb Haemost
JT  - Journal of thrombosis and haemostasis : JTH
JID - 101170508
RN  - 0 (Cardiolipins)
RN  - 0 (Polyphosphates)
RN  - 0 (Recombinant Proteins)
RN  - 9005-49-6 (Heparin)
RN  - EC 3.4.21.- (HABP2 protein, human)
RN  - EC 3.4.21.- (Serine Endopeptidases)
RN  - EC 3.4.21.21 (Factor VIIa)
RN  - EC 3.4.21.73 (Urokinase-Type Plasminogen Activator)
RN  - EC 3.4.24.27 (Thermolysin)
SB  - IM
CIN - J Thromb Haemost. 2012 May;10(5):857-8. PMID: 22394423
MH  - *Blood Coagulation/drug effects
MH  - Cardiolipins/metabolism
MH  - Enzyme Activation
MH  - Enzyme Stability
MH  - Factor VIIa/*metabolism
MH  - Fibrinolysis
MH  - Heparin/pharmacology
MH  - Humans
MH  - Kinetics
MH  - Polyphosphates/metabolism
MH  - Protein Denaturation
MH  - Recombinant Proteins/metabolism
MH  - Serine Endopeptidases/genetics/*metabolism
MH  - Substrate Specificity
MH  - Thermolysin/metabolism
MH  - Urokinase-Type Plasminogen Activator/metabolism
EDAT- 2012/01/13 06:00
MHDA- 2012/08/31 06:00
CRDT- 2012/01/13 06:00
PHST- 2012/01/13 06:00 [entrez]
PHST- 2012/01/13 06:00 [pubmed]
PHST- 2012/08/31 06:00 [medline]
AID - S1538-7836(22)06250-X [pii]
AID - 10.1111/j.1538-7836.2012.04619.x [doi]
PST - ppublish
SO  - J Thromb Haemost. 2012 May;10(5):859-66. doi: 10.1111/j.1538-7836.2012.04619.x.