A nonspecific, irreversible serine protease inhibitor
PMSF (Phenylmethanesulfonyl fluoride) is an irreversible serine/cysteine protease inhibitor and PMSF commonly used in the preparation of cell lysates [1,2]. phenylmethylsulfonyl fluoride (PMSF) is also an inhibitor of fatty acid amide hydrolase (FAAH) [3].
Aqueous preparations of PMSF become inactive toward proteases unless promptly brought into contact with protease. Inactivation of PMSF increases with increased pH and temperature. Half-lives of the inhibitor at 25°C are approximately 110,55, and 35 min at pH 7.0, 7.5, and 8.0, respectively. At pH 8, 100 μM PMSF is almost completely inactivated within 1 hr at 25°C or within 22 hr at 4°C. Stock solutions of PMSF in 100% isopropanol are stable at 25°C for months if not longer. Reactivation of PMSF-inhibited chymotrypsin did not occur within 1 week at 25°C at pH 7.0 [4]. PMSF inhibits the acylation of the inositol residue of GPI intermediates in bloodstream form T. brucei. PMSF inhihits the formntion of glycolipid C but does not inhibit fatty acid remodeling in vitro. PMSF inhihits GPI acylation and ethanolamine phosphatp addition in procyclic trypanosomes but not in Hela cells [5].
PMSF has a notable antinociceptive effect at doses 120 and 300 mg/kg. The dose of (60 mg/kg, i.p.) PMSF was considered as a sub-antinociceptive dose. A sub-antinociceptive dose (60 mg/kg) of PMSF could reduce tolerance in both acute and chronic methods of administration. However, alleviation of dependence and suppression of oxidative stress markers occurred in the chronic administration of PMSF [3].
参考文献:
[1]. Tunlid A, Jansson S. Proteases and their involvement in the infection and immobilization of nematodes by the nematophagous fungus Arthrobotrys oligospora[J]. Applied and Environmental Microbiology, 1991, 57(10): 2868-2872.
[2]. Rajakumar A, Brandon H M, Daftary A, et al. Evidence for the functional activity of hypoxia-inducible transcription factors overexpressed in preeclamptic placentae[J]. Placenta, 2004, 25(10): 763-769.
[3]. Akbarabadi E A, Vardanjani H R, Molavinia S, et al. PMSF Attenuates Morphine Antinociceptive Tolerance and Dependence in Mice: Its Association with the Oxidative Stress Suppression[J]. Iranian Journal of Pharmaceutical Research: IJPR, 2021, 20(3): 300.
[4]. James G T. Inactivation of the protease inhibitor phenylmethylsulfonyl fluoride in buffers[J]. Analytical biochemistry, 1978, 86(2): 574-579.
[5]. Güther M L, Masterson W J, Ferguson M A. The effects of phenylmethylsulfonyl fluoride on inositol-acylation and fatty acid remodeling in African trypanosomes[J]. Journal of Biological Chemistry, 1994, 269(28): 18694-18701.
PMSF（苯甲磺酰氟）是一种不可逆的丝氨酸/半胱氨酸蛋白酶抑制剂，PMSF 常用于制备细胞裂解物[1,2]。苯甲基磺酰氟 (PMSF) 也是脂肪酸酰胺水解酶 (FAAH) 的抑制剂[3]。
除非立即与蛋白酶接触，否则 PMSF 的水性制剂对蛋白酶失去活性。 PMSF 的失活随着 pH 值和温度的升高而增加。在 25°C 和 pH 7.0、7.5 和 8.0 时，抑制剂的半衰期分别约为 110,55 和 35 分钟。在 pH 8 时，100 μM PMSF 在 25°C 下 1 小时内或在 4°C 下 22 小时内几乎完全失活。 PMSF 的 100% 异丙醇储备溶液在 25°C 下可稳定数月甚至更长。在 25°C 和 pH 7.0 下 1 周内，PMSF 抑制的胰凝乳蛋白酶没有发生再激活 [4]。 PMSF 抑制血流中 GPI 中间体肌醇残基的酰化形成布氏锥虫。 PMSF 抑制糖脂 C 的形成，但不抑制体外脂肪酸重塑。 PMSF 抑制前环锥虫中的 GPI 酰化和乙醇胺磷酸化添加，但不抑制 Hela 细胞[5]。
PMSF 在剂量为 120 和 300 mg/kg 时具有显着的镇痛作用。 (60 mg/kg, i.p.) PMSF 的剂量被认为是亚镇痛剂量。亚镇痛剂量 (60 mg/kg) 的 PMSF 可以降低急性和慢性给药方法的耐受性。然而，长期服用 PMSF 可减轻依赖性并抑制氧化应激标志物 [3]。
| Animal experiment [1]: | |
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Animal models |
Adult male mice |
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Preparation Method |
This assay measures analgesic activity. All animals were placed on a 55 ± 1 °C hot plate, and the latency time to either jumping or licking was recorded. The cut-off time was set as 15 s to protect mice from any damage. The antinociceptive effects of a single dose of morphine (10 mg/kg s.c.) and PMSF (60, 120, and 300 mg/kg) were determined. |
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Dosage form |
60, 120, and 300 mg/kg, i.p. |
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Applications |
The results suggest that the administration of the PMSF doses (120 and 300 mg/kg, i.p.) led to significant changes in the analgesic latency time, while administration with PMSF (60 mg/kg i.p.) had no significant difference. |
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参考文献: [1]: Akbarabadi E A, Vardanjani H R, Molavinia S, et al. PMSF Attenuates Morphine Antinociceptive Tolerance and Dependence in Mice: Its Association with the Oxidative Stress Suppression[J]. Iranian Journal of Pharmaceutical Research: IJPR, 2021, 20(3): 300. |
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