Design of cancer immunostimulant peptide by designable proteomix

通过可设计蛋白质组设计癌症免疫刺激肽

• 批准号: 15360439
• 负责人: HONDA Hiroyuki
• 金额: $ 9.54万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15360439/
• 关键词: Peotide; Proteomix; Cancer; Conputational science; Peptide chip; Exhaustive; Immunostimulant; HLA; バイオインフォマティクス; HSP70; Fas ligand; Angiotensin II

Peptides, which are small biocompatible molecules with simple structure as compared to proteins, have shown various applications functioning as an antimicrobial molecule, inhibitory drug, targeting molecule, growth factor, and nano-material for biocompatible matrixes. Peptide arrays, developed as the SPOT method by R.Frank, have shown great efficiency in high-throughput screening of peptides for biomedical and pharmaceutical researches. In the SPOT method, activated amino acid monomers are spotted on support, such as cellulose membrane, to form a spatially addressed peptide library with the objective peptide length and order. In this study, we demonstrate the efficiency of our screening strategy in which a peptide array was used for direct peptide-cell interaction. In addition, the data obtained were also analyzed by computational science to establish the new method for functional peptide design, "designable proteomics". Consequently, the following results were obtained.1)Screening of … More newry functional peptidesPeptide arrays covering the Ang II receptor type 1 sequence were prepared, and the peptide (VVIVIY) within the first transmembrane region exhibited the highest affinity to Ang II. The synthesized soluble VVIVIY peptide bad an 84% inhibitory effect on Ang II-induced aorta contration. In addition, we investigated the efficiency of the use of peptide array for direct cell assay, without cleavage of peptides, to design novel tumor growth-inhibitory peptides. From human Fas antigen ligand sequence, we found a novel 5-mer peptide domain (CNNLP) that strongly inhibits the growth of cancer cells by interaction with cell surface.2)Analysis of peptide chip data by computational science to establish "designable proteomics"2-1)HMM model construction for peptide screeningHidden Markov model was constructed for screening of peptide combined with MHC class II molecule. The prediction acuracy of the model constructed was more than 90%.2-2)Design of cell death inducing peptideThe effect of substitution of one peptide in the sequence of CNNLP was investigated by peptide chip cell assay system. From computational analysis of the data, the following rule was obtained to design the functional peptide ; 3 amino acid residues are important for the function and N-terminal amino acid should be small. Especially, threonin or valine is effective.2-3)Exploring of lipases with inverted enantioselectivity for a substrateWe explored lipases with inverted enantioselectivity from the wild-type bacterial lipase, which is originally selective for (S)-configuration of the substrate. Computational tool constructed by us predicted a rule indicating that "size at position L167," among 4 positions(L17,F119,L167,and L266) in the substrate binding core region, is the most influential factor. Based on the guidelines obtained, we found two engineered novel variants, namely FIGV(L17F,F119I,L167G,and L266V) and FFGI(L17F,L167G,and L266I).3)Exploring of cancer vaccine peptideFrom the sequence of HSP70,the exploring of candidates of cancer vaccine peptide was investigated. HSP70 was one of the cancer specific proteins and a part of protein sequence is possible to become cancer vaccine peptide. From the assay of peptide chip spotting HSP70 sequence, more than 10 candidate peptide was obtained. Less

与蛋白质相比，肽是具有简单结构的小生物相容性分子，其已经显示出作为抗微生物分子、抑制药物、靶向分子、生长因子和用于生物相容性基质的纳米材料的各种应用。由R.Frank开发的肽阵列作为SPOT方法，在生物医学和药学研究的肽的高通量筛选中显示出极大的效率。在SPOT方法中，将活化的氨基酸单体点样在支持物如纤维素膜上，以形成具有目标肽长度和顺序的空间寻址肽文库。在这项研究中，我们证明了我们的筛选策略的效率，其中肽阵列用于直接肽-细胞相互作用。此外，还利用计算科学对所得数据进行了分析，建立了功能肽设计的新方法--“可设计蛋白质组学”。因此，获得了以下结果：1）筛选 ...更多信息 新功能肽制备了覆盖Ang Ⅱ 1型受体序列的肽阵列，其中第一跨膜区的肽（VVIVIY）对Ang Ⅱ表现出最高的亲和力。合成的可溶性VVIVIY肽对Ang Ⅱ诱导的主动脉收缩有84%的抑制作用。此外，我们研究了使用肽阵列进行直接细胞测定的效率，而不切割肽，以设计新的肿瘤生长抑制肽。从人Fas抗原配体序列中，我们发现了一个新的5聚体肽结构域（CNNLP），它通过与细胞表面的相互作用强烈抑制癌细胞的生长。2）通过计算科学分析肽芯片数据，建立“可设计的蛋白质组学”2-1）用于肽筛选的隐马尔可夫模型构建隐马尔可夫模型用于筛选与MHC II类分子结合的肽。2 -2）细胞死亡诱导肽的设计利用肽芯片细胞分析系统研究了CNNLP序列中一个肽的取代对CNNLP的影响。从数据的计算分析，得到以下规则来设计功能肽; 3个氨基酸残基对于功能是重要的，并且N-末端氨基酸应该小。2 -3）对底物具有反向对映选择性的脂肪酶的探索我们从野生型细菌脂肪酶中探索了具有反向对映选择性的脂肪酶，该脂肪酶最初对底物的（S）-构型具有选择性。我们构建的计算工具预测了一个规则，表明底物结合核心区的4个位置（L17、F119、L167和L266）中“L167位置的大小”是最有影响的因素。在此基础上，我们发现了两个新的工程变体，即FIGV（L17 F，F119 I，L167 G和L266 V）和FFGI（L17 F，L167 G和L266 I）。3）癌症疫苗肽的探索从HSP 70序列出发，探索癌症疫苗肽的候选肽。HSP 70是一种肿瘤特异性蛋白，其部分序列有可能成为肿瘤疫苗肽。通过肽芯片点样HSP 70序列，筛选出10多个候选肽段。少

项目成果

ナノ バイオテクノロジーの最前線

纳米生物科技前沿

• 发表时间: 2003
• 作者: 本多裕之;加藤竜司
• 通讯作者: 加藤竜司

Informatics supporting the research field of nanotechnology(in Japanese)

支持纳米技术研究领域的信息学（日语）

• 发表时间: 2003
• 期刊: Front Line of Nanobiotechnology(ed., M.Ueda)(CMC Shuppan)
• 作者: H.Honda;R.Kato
• 通讯作者: R.Kato

Direct cell assay on peptide array is effective for functional peptide design

肽阵列上的直接细胞测定对于功能肽设计是有效的

• 期刊: Biochemical and Biophysical Research Communications (submitted)
• 作者: R.Kato;Y.Okuno;C.Kaga;M.Kunimatsu;T.Kobayashi;H.Honda
• 通讯作者: H.Honda

Ryuji Kato, Hideki Noguchi.Hiroyuki Honda, Takeshi Kobayashi: "Hidden Markov model-based approach as the first screening of binding peptides that interact with MHC class II molecules"Enzyme and Microbial Technology. 33. 472-481 (2003)

Ryuji Kato、Hideki Noguchi、Hiroyuki Honda、Takeshi Kobayashi：“基于隐马尔可夫模型的方法作为与 MHC II 类分子相互作用的结合肽的首次筛选”酶和微生物技术。

Angiotensin II inhibitory peptide found in the receptor sequence using peptide array

使用肽阵列在受体序列中发现血管紧张素 II 抑制肽

• 发表时间: 2004
• 期刊: Biochemical and Biophysical Research Communications 315
• 作者: Ryuji Kato;Mitoshi Kunimatsu;Takeshi Kobayashi;Hiroyuki Honda
• 通讯作者: Hiroyuki Honda