title: A novel serine protease with human fibrino(geno)lytic activities from Artocarpus heterophyllus latex.
abstract: A protease was isolated and purified from Artocarpus heterophyllus (jackfruit) latex and designated as a 48-kDa antimicrobial protease (AMP48) in a previous publication. In this work, the enzyme was characterized for more biochemical and medicinal properties. Enzyme activity of AMP48 was strongly inhibited by phenylmethanesulfonyl fluoride and soybean trypsin inhibitor, indicating that the enzyme was a plant serine protease. The N-terminal amino acid sequences (A-Q-E-G-G-K-D-D-D-G-G) of AMP48 had no sequence similarity matches with any sequence databases of BLAST search and other plant serine protease. The secondary structure of this enzyme was composed of high α-helix (51%) and low β-sheet (9%). AMP48 had fibrinogenolytic activity with maximal activity between 55 and 60°C at pH 8. The enzyme efficiently hydrolyzed α followed by partially hydrolyzed β and γ subunits of human fibrinogen. In addition, the fibrinolytic activity was observed through the degradation products by SDS-PAGE and emphasized its activity by monitoring the alteration of secondary structure of fibrin clot after enzyme digestion using ATR-FTIR spectroscopy. This study presented the potential role to use AMP48 as antithrombotic for treatment thromboembolic disorders such as strokes, pulmonary emboli and deep vein thrombosis.
authors.affiliation: Institute of Science, Suranaree University of Technology, Nakhon Ratchasima, Thailand. jaruwan_siritape@yahoo.com
authors.forename: Jaruwan
authors.initials: J
authors.lastname: Siritapetawee
authors.forename: Kanjana
authors.initials: K
authors.lastname: Thumanu
authors.forename: Punchapat
authors.initials: P
authors.lastname: Sojikul
authors.forename: Sompong
authors.initials: S
authors.lastname: Thammasirirak
mesh_terms: D000595:Amino Acid Sequence; D031622:Artocarpus / Q000737:chemistry*; D005340:Fibrinogen / Q000737:chemistry*; D005343:Fibrinolytic Agents / Q000737:chemistry* / Q000302:isolation & purification; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007840:Latex / Q000737:chemistry*; D008969:Molecular Sequence Data; D010664:Phenylmethylsulfonyl Fluoride / Q000737:chemistry; D010940:Plant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D017433:Protein Structure, Secondary; D021122:Protein Subunits / Q000737:chemistry*; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature; D014361:Trypsin Inhibitors / Q000737:chemistry
chemical_list: D005343:Fibrinolytic Agents; D007840:Latex; D010940:Plant Proteins; D021122:Protein Subunits; D014361:Trypsin Inhibitors; D010664:Phenylmethylsulfonyl Fluoride; D005340:Fibrinogen; D012697:Serine Endopeptidases