Expression of the fgl2 and Its Protein Product (Prothrombinase) in Tissues During Murine Hepatitis Virus Strain-3 (MHV-3) Infection

Ding, Jin; Ning, Qin; Liu, M. F.; Lai, Alexander; Peltekian, Kevork M.; Fung, Laisum; Holloway, C. J.; Yeger, Herman; Phillips, M. James; Levy, Gary

doi:10.1007/978-1-4615-5331-1_79

Cited by 43 publications

(48 citation statements)

References 13 publications

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“…Several lines of evidence indicate fgl2 exhibits coagulation activity. First, following MHV-3 infection, mRNA transcripts of fgl2 and fgl2 protein can be detected predominantly in liver reticuloendothelial cells, followed by fibrin deposition and widespread hepatic necrosis (13). We have recently reported a similar relationship of fgl2 to fulminant hepatitis B virus infection (1).…”

mentioning

confidence: 58%

“…Homogeneity of purified soluble fgl2 protein was evaluated by SDS-PAGE and confirmed by Western blot probed with anti-fgl2 Ab as previously described (13). Proteins were stained directly using Coomassie brilliant blue or were transferred to nitrocellulose, and then were probed using polyclonal rabbit anti-mouse fgl2 IgG as the primary Ab.…”

Section: Expression Of Fgl2 Proteinmentioning

confidence: 99%

“…Our laboratory has previously demonstrated the pivotal role of a unique direct prothrombinase, fgl2, in the pathogenesis of fulminant viral hepatitis secondary to murine hepatitis virus strain-3 (MHV-3) 3 infection (11)(12)(13). The gene fgl2 was originally cloned from CTLs, and the encoded protein was named as a fibrinogenlike protein due to homology to the carboxyl terminus of the ␤-and ␥-chains of fibrinogen (14).…”

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confidence: 99%

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Kinetic Analysis of a Unique Direct Prothrombinase, fgl2, and Identification of a Serine Residue Critical for the Prothrombinase Activity

Chan

Fung

et al. 2002

The Journal of Immunology

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fgl2 prothrombinase, by its ability to generate thrombin, has been shown to be pivotal to the pathogenesis of viral-induced hepatitis, cytokine-induced fetal loss syndrome, and xeno- and allograft rejection. In this study, the molecular basis of fgl2 prothrombinase activity was examined in detail. Purified fgl2 protein generated in a baculovirus expression system had no measurable prothrombinase activity, whereas the activity was restored when the purified protein was reconstituted into phosphatidyl-l-serine-containing vesicles. Reconstituted fgl2 catalyzed the cleavage of human prothrombin to thrombin with kinetics consistent with a first order reaction, with an apparent Vmax value of 6 mol/min/mol fgl2 and an apparent Km value for prothrombin of 8.3 μM. The catalytic activity was totally dependent on calcium, and factor Va (500 nM) enhanced the catalytic efficiency of fgl2 by increasing the apparent Vmax value to 3670 mol/min/mol fgl2 and decreasing the apparent Km value for prothrombin to 7.2 μM. By a combination of site-directed mutagenesis and production of truncated proteins, it was clearly shown that residue Ser89 was critical for the prothrombinase activity of fgl2. Furthermore, fgl2 prothrombinase activity was not inhibited by antithrombin III, soybean trypsin inhibitor, 4-aminobenzamidine, aprotinin, or phenylmethylsulfonyl fluoride, whereas diisopropylfluorophosphate completely abrogated the activity. In this work we provide direct evidence that fgl2 cleaves prothrombin to thrombin consistent with serine protease activity and requires calcium, phospholipids, and factor Va for its full activity.

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mentioning

confidence: 58%

Section: Expression Of Fgl2 Proteinmentioning

confidence: 99%

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confidence: 99%

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Kinetic Analysis of a Unique Direct Prothrombinase, fgl2, and Identification of a Serine Residue Critical for the Prothrombinase Activity

Chan

Fung

et al. 2002

The Journal of Immunology

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“…For fibrin detection, a rabbit-anti-fibrinogen Ab (DakoCytomation) was used. This reagent is known to react with fibrinogen and fibrin in mouse and human tissues (1,9). The technique used for detection of fibrin was the standard avidin-biotin complex (ABC) method as previously described (9).…”

Section: In Situ Hybridizationmentioning

confidence: 99%

“…The method used has been described previously (1). A digoxigenin-11-UTP (Dig-UTP) (Roche)-labeled cDNA probe was cut by EcoRI following subcloning of a 169-bp fragment of mfgl2 cDNA, representing nt 756 (ACTGTGACA.…”

Section: In Situ Hybridizationmentioning

confidence: 99%

Role of Fibrinogen-Like Protein 2 Prothrombinase/Fibroleukin in Experimental and Human Allograft Rejection

Ning

Sun

Han

et al. 2005

The Journal of Immunology

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Immune coagulation is a major contributor to the pathogenesis of xenograft rejection, viral-induced hepatocellular injury and cytokine-induced fetal loss syndrome. In this study, we investigated the contribution of the novel gene product, fibrinogen-like protein 2 (fgl2) prothrombinase, in mediating immune injury in experimental and human acute allograft rejection. Using a mouse heterotopic cardiac transplant model, mouse fgl2(mfgl2)/fibroleukin mRNA transcripts and protein were highly expressed in macrophages, CD4- and CD8-positive T lymphocytes, and endothelial cells in rejecting cardiac allografts in association with deposits of fibrin. Although mfgl2-deficient mice rejected allografts at similar rates to littermate controls, survival of grafts from mfgl2-deficient mice were prolonged and deposition of intravascular fibrin was diminished. Treatment of wild-type mice with a neutralizing anti-fgl2 Ab ameliorated histological evidence for allorejection and intravascular fibrin deposition, and resulted in an increase in graft survival. To address further the relevance of fgl2 in acute allograft rejection, we examined kidney biopsies from patients who had undergone renal transplantation. Human fgl2 mRNA transcripts and protein were markedly expressed mainly in renal tubule cells, infiltrating lymphoid cells including macrophages, CD8+ T cells, mature B cells (plasma cells), and endothelial cells. Dual staining showed fibrin deposition was localized mainly to blood vessels, in the glomerulus and interstitium and the lumen of tubules, and occurred in association with human fgl2 expression. These data collectively suggest that fgl2 accounts for the fibrin deposition seen in both experimental and human allograft rejection and provide a rationale for targeting fgl2 as adjunctive therapy to treat allograft rejection.

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Adenovirus‐mediated artificial microRNA against human fibrinogen like protein 2 inhibits hepatocellular carcinoma growth

Wang

Liu

et al. 2016

The Journal of Gene Medicine

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Expression of the fgl2 and Its Protein Product (Prothrombinase) in Tissues During Murine Hepatitis Virus Strain-3 (MHV-3) Infection

Cited by 43 publications

References 13 publications

Kinetic Analysis of a Unique Direct Prothrombinase, fgl2, and Identification of a Serine Residue Critical for the Prothrombinase Activity

Kinetic Analysis of a Unique Direct Prothrombinase, fgl2, and Identification of a Serine Residue Critical for the Prothrombinase Activity

Role of Fibrinogen-Like Protein 2 Prothrombinase/Fibroleukin in Experimental and Human Allograft Rejection

Adenovirus‐mediated artificial microRNA against human fibrinogen like protein 2 inhibits hepatocellular carcinoma growth

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