基于马豆种子 Bowman-Birk 抑制剂结合环的小分子胰蛋白酶抑制剂的分子工程。

Molecular engineering of a small trypsin inhibitor based on the binding loop of horsegram seed Bowman-Birk inhibitor.

机构信息

Department of Protein Chemistry and Technology, Central Food Technological Research Institute, Council of Scientific and Industrial Research, Mysore, India.

出版信息

J Enzyme Inhib Med Chem. 2011 Aug;26(4):553-60. doi: 10.3109/14756366.2010.536158. Epub 2010 Dec 20.

DOI:10.3109/14756366.2010.536158

PMID:21171897

Abstract

CONTEXT

The Bowman-Birk inhibitors (BBIs) are currently investigated with renewed interest due to their therapeutic properties in cancer and other inflammatory disease treatment. The molecular mass of the BBI is a limitation, as sufficient amounts of the inhibitor do not reach the organs outside the gastrointestinal tract when administered orally.

METHOD

The anti-tryptic domain of HGI-III of horsegram (Dolichos biflorus) was cloned using the vector pET-20b (+) and expressed in E. coli BL21 (DE3) pLysS.

RESULTS

Kinetic analysis of this anti-tryptic peptide (recombinant trypsin inhibitory domain (rTID)) reveals that it is a potent inhibitor of trypsin and human tryptase. The K(i) (3.2 ± 0.17 × 10(-8) M) establishes a very high affinity to bovine trypsin. rTID inhibited human lung tryptase (IC(50) 3.78 ± 0.23 × 10(-7) M). The rTID is resistant to the digestive enzymes found in humans and animals.

CONCLUSION

These properties propagate further research on the use of rTID as a therapeutic for cancer and other related inflammatory diseases.

摘要

背景

由于在癌症和其他炎症性疾病治疗方面的治疗特性，目前人们对 Bowman-Birk 抑制剂（BBIs）重新产生了兴趣。BBI 的分子质量是一个限制因素，因为当口服给药时，足够量的抑制剂不会到达胃肠道以外的器官。

方法

使用载体 pET-20b（+）克隆马豆（Dolichos biflorus）的 HGI-III 的抗胰蛋白酶结构域，并在 E. coli BL21（DE3）pLysS 中表达。

结果

该抗胰蛋白酶肽（重组胰蛋白酶抑制结构域（rTID））的动力学分析表明，它是胰蛋白酶和人胰蛋白酶的有效抑制剂。该 K(i)（3.2±0.17×10(-8)M）与人牛胰蛋白酶建立了非常高的亲和力。rTID 抑制人肺胰蛋白酶（IC(50)3.78±0.23×10(-7)M）。rTID 对人和动物体内发现的消化酶具有抗性。