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A Protein from Aloe vera that Inhibits the Cleavage of Human Fibrin(ogen) by Plasmin

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Abstract

A protease inhibitor protein with the molecular mass of 11,804.931 Da (analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry) was isolated from Aloe vera leaf gel and designated as AVPI-12. The isoelectric point of the protein is about 7.43. The first ten amino acid sequence from the N-terminal was found to be R–D–W–A–E–P–N–D–G–Y, which did not match other protease inhibitors in database searches and other publications, indicating AVPI-12 is a novel protease inhibitor. The band protein of AVPI-12 migrated further on nonreducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) than reducing SDS-PAGE. This result indicated that the molecule of AVPI-12 did not contain interchain disulfide bonds, but appeared to have intrachain disulfide bonds instead. AVPI-12 strongly resisted digestion by the serine proteases human plasmin and bovine trypsin. The protein could protect the γ-subunit of human fibrinogen from plasmin and trypsin digestion, similar to the natural plasma serine protease inhibitor α2-macroglobulin. The protein also could protect the γ-subunit of fibrinogen from the cysteine protease papain. AVPI-12 also exhibited dose-dependent inhibition of the fibrinogenolytic activity of plasmin, similar to α2-macroglobulin. The fibrinolytic inhibitory activity of AVPI-12 and the small-angle X-ray scattering showed that the protein could protect human fibrin clot from complete degradation by plasmin. The inhibition of the fibrinogenolytic and fibrinolytic activities of plasmin by AVPI-12 suggests that the inhibitor has potential for use in antifibrinolytic treatment.

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Acknowledgments

This research was supported by the Suranaree University of Technology Research Grant (SUT1-102-55-12-34). We also thank the Synchrotron Light Research Institute (Public Organization), Nakhon Ratchasima, Thailand for providing facilities to do the SAXS experiment at BL-2.2 and analyses.

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Authors and Affiliations

  1. School of Biochemistry, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima, Thailand

    Jaruwan Siritapetawee

  2. Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok, Thailand

    Punchapat Sojikul

  3. Synchrotron Light Research Institute (Public Organization), Nakhon Ratchasima, Thailand

    Siriwat Soontaranon & Wanwisa Limphirat

  4. Protein and Proteomics Research Center for Commercial and Industrial Purposes (ProCCI), Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand

    Sompong Thammasirirak

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  1. Jaruwan Siritapetawee
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  2. Punchapat Sojikul
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  4. Wanwisa Limphirat
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  5. Sompong Thammasirirak
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Correspondence to Jaruwan Siritapetawee.

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Siritapetawee, J., Sojikul, P., Soontaranon, S. et al. A Protein from Aloe vera that Inhibits the Cleavage of Human Fibrin(ogen) by Plasmin. Appl Biochem Biotechnol 170, 2034–2045 (2013). https://doi.org/10.1007/s12010-013-0356-2

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  • DOI: https://doi.org/10.1007/s12010-013-0356-2

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