Engineered cystine knot peptides that bind alphavbeta3, alphavbeta5, and alpha5beta1 integrins with low-nanomolar affinity.
Engineered cystine knot peptides that bind alphavbeta3, alphavbeta5, and alpha5beta1 integrins with low-nanomolar affinity.
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DOI：
10.1002/prot.22441
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发表时间：
2009-11-01
期刊：
影响因子：
2.9
通讯作者：
Cochran JR
中科院分区：
文献类型：
--
作者：
Kimura RH;Levin AM;Cochran FV;Cochran JR
对癌症治疗和诊断的应用靶向细胞表面整合素受体的迫切需求。一类新的整联蛋白结合剂。包含Arg-Gly-Asp（RGD）整合素的结合基序和随机侧翼残差以高通量方式筛选。显示出与在U87MG胶质母细胞瘤细胞表面表达的整联蛋白受体结合的自然配体结合，具有半最大抑制浓度（IC50）值10-30 nm的受体特异性表明，工程性的肽与高亲和力的αVβ3和αVβ5整合素结合，我们还发现了与αVβ3，αVβ5和α5β1整合素的高亲和力结合的肽。由于所有这三个受体都可以在肿瘤上共表达，因此我们表明了工程化的辣椒细胞会抑制肿瘤细胞根据其整合素的结合性，这些数据粘附于细胞外基质蛋白玻璃蛋白，在某些情况下，这些数据将EETI-II验证为蛋白质工程的脚手架，并突出了独特的整合素结合的Pepperides的发展在癌症中的应用。
There is a critical need for compounds that target cell surface integrin receptors for applications in cancer therapy and diagnosis. We used directed evolution to engineer the Ecballium elaterium trypsin inhibitor (EETI-II), a knottin peptide from the squash family of protease inhibitors, as a new class of integrin-binding agents. We generated yeast-displayed libraries of EETI-II by substituting its 6-amino acid trypsin binding loop with 11-amino acid loops containing the Arg-Gly-Asp (RGD) integrin binding motif and randomized flanking residues. These libraries were screened in a high-throughput manner by fluorescence-activated cell sorting to identify mutants that bound to αvβ3 integrin. Select peptides were synthesized and were shown to compete for natural ligand binding to integrin receptors expressed on the surface of U87MG glioblastoma cells with half-maximal inhibitory concentration (IC50) values of 10-30 nM. Receptor specificity assays demonstrated that engineered knottin peptides bind to both αvβ3 and αvβ5 integrins with high affinity. Interestingly, we also discovered a peptide that binds with high affinity to αvβ3, αvβ5, and α5β1 integrins. This finding has important clinical implications since all three of these receptors can be co-expressed on tumors. In addition, we showed that engineered knottin peptides inhibit tumor cell adhesion to the extracellular matrix protein vitronectin, and in some cases fibronectin, depending on their integrin binding specificity. Collectively, these data validate EETI-II as a scaffold for protein engineering, and highlight the development of unique integrin-binding peptides with potential for translational applications in cancer.
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影响因子：
6.8
作者：
Gai, S. Annie;Wittrup, K. Dane
通讯作者：
Wittrup, K. Dane
影响因子：
5.6
作者：
Gilbreth, Ryan N.;Esaki, Kaori;Koide, Shohei
通讯作者：
Koide, Shohei
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：
56.9
作者：
BROOKS, PC;CLARK, RAF;CHERESH, DA
通讯作者：
CHERESH, DA
影响因子：
4.8
作者：
Hilpert, K;Wessner, H;Höhne, W
通讯作者：
Höhne, W