Combinatorial optimization of cystine-knot peptides towards high-affinity inhibitors of human matriptase-1.
Combinatorial optimization of cystine-knot peptides towards high-affinity inhibitors of human matriptase-1.
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DOI：
10.1371/journal.pone.0076956
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Kolmar H
中科院分区：
文献类型：
--
作者：
Glotzbach B;Reinwarth M;Weber N;Fabritz S;Tomaszowski M;Fittler H;Christmann A;Avrutina O;Kolmar H
半胱氨酸结小蛋白定义了一类具有数千个自然成员的生物活性分子。它们的同名基序包括一个由六个二硫键连接的半胱氨酸残基形成的刚性结构核心，这解释了它对热或蛋白质降解的特殊稳定性。由于它们在二硫键连接的环中表现出显著的序列耐受性，这些分子被认为是基于多肽的药物的有前途的框架。天然开链半胱氨酸结胰酶抑制剂MCoTI(苦瓜胰酶抑制剂)和SOTI(Spinacia Lenacea Trypsin Inhibitor)家族是合成Mattritase-1抑制剂的起点，Mattritase-1是一种可能与癌症和关节炎治疗相关的II型跨膜丝氨酸蛋白酶。酵母表面展示的小蛋白文库被用来选择独特和有效的Mattritase-1抑制剂。为此，应用了基于知识的文库设计，该文库利用了关于胱氨酸结肽的结合和折叠行为的详细信息。我们鉴定了5个抑制剂变体，其中4个属于MCoTI家族，1个属于SOTI家族，它们被化学合成并被氧化折叠成生物活性构象。酶分析显示，所有候选者的抑制常数都在低纳摩尔范围内。鉴定出一种对胰酶和基质裂解酶-1具有反向选择性的亚纳摩尔结合剂(KI = 0.83nM)。
Cystine-knot miniproteins define a class of bioactive molecules with several thousand natural members. Their eponymous motif comprises a rigid structured core formed by six disulfide-connected cysteine residues, which accounts for its exceptional stability towards thermic or proteolytic degradation. Since they display a remarkable sequence tolerance within their disulfide-connected loops, these molecules are considered promising frameworks for peptide-based pharmaceuticals. Natural open-chain cystine-knot trypsin inhibitors of the MCoTI (Momordica cochinchinensis trypsin inhibitor) and SOTI (Spinacia oleracea trypsin inhibitor) families served as starting points for the generation of inhibitors of matriptase-1, a type II transmembrane serine protease with possible clinical relevance in cancer and arthritic therapy. Yeast surface-displayed libraries of miniproteins were used to select unique and potent matriptase-1 inhibitors. To this end, a knowledge-based library design was applied that makes use of detailed information on binding and folding behavior of cystine-knot peptides. Five inhibitor variants, four of the MCoTI family and one of the SOTI family, were identified, chemically synthesized and oxidatively folded towards the bioactive conformation. Enzyme assays revealed inhibition constants in the low nanomolar range for all candidates. One subnanomolar binder (Ki = 0.83 nM) with an inverted selectivity towards trypsin and matriptase-1 was identified.
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DOI：
10.1093/protein/12.9.797
发表时间：
1999-09-01
期刊：
PROTEIN ENGINEERING
影响因子：
--
作者：
Christmann, A;Walter, K;Kolmar, H
通讯作者：
Kolmar, H
影响因子：
3.2
作者：
Avrutina, Olga;Schmoldt, Hans-Ulrich;Kolmar, Harald
通讯作者：
Kolmar, Harald
影响因子：
4
作者：
Getz, Jennifer A.;Rice, Jeffrey J.;Daugherty, Patrick S.
通讯作者：
Daugherty, Patrick S.
影响因子：
3.5
作者：
Désilets, A;Longpré, JM;Leduc, R
通讯作者：
Leduc, R
影响因子：
2.8
作者：
Craik, DJ;Daly, NL;Waine, C
通讯作者：
Waine, C