Characterization of a Metalloenzyme from a Wild Mushroom, Tricholoma saponaceum

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概要

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• Two kinds of metalloendopeptidases from the fruiting bodies of Tricholoma saponaceum (TSMEP1 and TSMEP2) have been purified, and TSMEP1 has been characterized based on their fibrinolytic activity. The enzymes have the same N-terminal amino acid sequence, Ala-Leu-Tyr-Val-Gly-X-Ser-Pro-X-Gln-Gln-Ser-Leu-Leu-Val, but slightly different molecular weights of 18,147 and 17,947,as measured by matrix assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry. The N-terminal sequence do not match with any known protein or open reading frame. TSMEP1 hydrolyzes fibrinogen as well as fibrin, but does not show any proteolytic activity for other blood proteins such as thrombin, human albumin, human IgG, hemoglobin, or urokinase. The enzyme hydrolyzes both Aα and Bβ subunits of human fibrinogen with equal efficiency but didn't show any reactivity for the γ form of human fibrinogen. The enzymatic activity is strongly inhibited by EDTA and 1,10-phenanthroline, indicating that the enzymes are metalloproteases. No inhibition was found with phenylmethylsulfonyl fluoride (PMSF), L-trans-epoxysuccinyl leucylamido-(4-guanidino)-butane (E-64), pepstatin and 2-mercaptoethanol. The activity of the purified enzyme was increased by Mg^<2+>, Fe^<2+>, Zn^<2+>, and Co^<2+>, and slightly decreased by Ca^<2+>, but the enzyme activity was dramatically decreased by Cu^<2+>, and totally inhibited by Hg^<2+>. It has broad substrate specificity for synthetic peptides, and keep the high activity from pH 7.5 to 9,suggesting that the purified enzyme was a basic protease. The enzyme was stable up to 30℃ and the maximum fibrinolytic activity was at 55℃.

• 社団法人日本農芸化学会の論文
• 2001-02-23

著者

• Kim Yang

  Division Of Infectious Diseases Asan Medical Center

• Kim Jun-ho

  Department Of Chemistry Sangji University

• Kim Yang

  Division Of Molecular Life Science And Center For Cell Signaling Research Ewha Womans University

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• Characterization of a Metalloenzyme from a Wild Mushroom, Tricholoma saponaceum
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