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首頁> 美國衛(wèi)生研究院文獻(xiàn)>ISRN Toxicology >Bmoo FIBMP-I: A New Fibrinogenolytic Metalloproteinase from Bothrops moojeni Snake Venom
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Bmoo FIBMP-I: A New Fibrinogenolytic Metalloproteinase from Bothrops moojeni Snake Venom

機(jī)譯：Bmoo FIBMP-I：來自Bothrops moojeni蛇毒的新型纖維蛋白原分解金屬蛋白酶

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摘要

A new fibrinogenolytic metalloproteinase (Bmoo FIBMP-I) was purified from Bothrops moojeni snake venom. This enzyme was isolated through a combination of three chromatographic steps (ion-exchange, molecular exclusion, and affinity chromatography). Analyses by reverse phase chromatography, followed by mass spectrometry, showed the presence of enzyme isoforms with average molecular mass of 22.8 kDa. The SDS-PAGE analyses showed a single chain of 27.6 kDa, in the presence and absence of reducing agent. The protein has a blocked N-terminal. One of the peptides obtained by enzymatic digestion of a reduced and S-alkylated isoform was completely sequenced by mass spectrometry (MS/MS). Bmoo FIBMP-I showed similarity with hemorrhagic factor and several metalloproteinases (MP). This enzyme degraded Aα-chain faster than the Bβ-chain and did not affect the γ-chain of bovine fibrinogen. The absence of proteolytic activity after treatment with EDTA, together with the observed molecular mass, led us to suggest that Bmoo FIBMP-I is a member of the P-I class of the snake venom MP family. Bmoo FIBMP-I showed pH-dependent proteolytic activity on azocasein, but was devoid of coagulant, defibrinating, or hemorrhagic activities. The kinetic parameters of proteolytic activity in azocasein were determined (V max = 0.4596 Uh−1nmol−1 ± 0.1031 and K m = 14.59 mg/mL ± 4.610).
機(jī)譯：從Bothrops moojeni蛇毒中純化出一種新的纖維蛋白原分解金屬蛋白酶（Bmoo FIBMP-I）。通過三個(gè)色譜步驟（離子交換，分子排阻和親和色譜）的組合來分離該酶。通過反相色譜分析，然后進(jìn)行質(zhì)譜分析，表明存在平均分子量為22.8 kDa的酶同工型。 SDS-PAGE分析顯示在存在和不存在還原劑的情況下，單鏈為27.6 kDa。該蛋白質(zhì)的N末端被封閉。通過質(zhì)譜（MS / MS）對(duì)通過還原和還原的S-烷基化同工型進(jìn)行酶消化而獲得的一種肽進(jìn)行了完全測(cè)序。 Bmoo FIBMP-1與出血性因子和幾種金屬蛋白酶（MP）相似。該酶降解Aα鏈的速度快于Bβ鏈，并且不影響牛纖維蛋白原的γ鏈。 EDTA處理后缺乏蛋白水解活性，再加上觀察到的分子量，使我們認(rèn)為Bmoo FIBMP-I是蛇毒MP家族P-I類的成員。 Bmoo FIBMP-1對(duì)偶氮酪蛋白顯示出pH依賴的蛋白水解活性，但沒有凝結(jié)，去纖維蛋白或出血活性。確定了偶氮酪蛋白中蛋白水解活性的動(dòng)力學(xué)參數(shù)（V max = 0.4596 Uh -1 nmol -1 ±0.1031和K m = 14.59 mg / mL±4.610）。

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