A phage display selected 7-mer peptide inhibitor of the Tannerella forsythia metalloprotease-like enzyme Karilysin can be truncated to Ser-Trp-Phe-Pro.
A phage display selected 7-mer peptide inhibitor of the Tannerella forsythia metalloprotease-like enzyme Karilysin can be truncated to Ser-Trp-Phe-Pro.
复制标题
噬菌体展示选择的连翘金属蛋白酶样酶 Karilysin 的 7 聚体肽抑制剂可被截短为 Ser-Trp-Phe-Pro。
DOI：
10.1371/journal.pone.0048537
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Riise E
中科院分区：
文献类型：
--
作者：
Skottrup PD;Sørensen G;Ksiazek M;Potempa J;Riise E
牙周炎是一种常见的慢性牙周病，是牙周病的重要组成部分。Karilysin是一种新发现的类金属蛋白酶，由T.艾西娅Karilysin调节宿主免疫应答，因此被认为是可能的药物靶标。在这项研究中，通过噬菌体展示选择了来自Karilysin（Kly 18）的催化结构域的肽。肽是线性的，具有低微摩尔结合亲和力。两个最好的结合物（肽14和肽15）共享共有序列XWFPXXXGGG。用与麦芽糖结合蛋白（MBP）的N-末端融合的肽15产生与MBP的肽15融合物。在大肠杆菌中表达了15-MBP。用纯化的融合蛋白验证Kly 18的特异性结合。化学合成的肽15（SWFPLRSGGG）可以抑制Kly 18和完整的Karilysin（Kly 48）的酶活性。此外，肽15可以减缓完整Kly 48到Kly 18的自动加工。WFP基序对抑制作用很重要，截短研究进一步证明N-末端丝氨酸对Kly 18抑制作用也很重要。SWFP肽具有在低微摩尔范围内的Ki值，其与完整肽相似15。结论SWFP是首次报道的Karilysin抑制剂，可作为Karilysin结构-功能研究的有用工具。
Tannerella forsythia is a gram-negative bacteria, which is strongly associated with the development of periodontal disease. Karilysin is a newly identified metalloprotease-like enzyme, that is secreted from T. forsythia. Karilysin modulates the host immune response and is therefore considered a likely drug target. In this study peptides were selected towards the catalytic domain from Karilysin (Kly18) by phage display. The peptides were linear with low micromolar binding affinities. The two best binders (peptide14 and peptide15), shared the consensus sequence XWFPXXXGGG. A peptide15 fusion with Maltose Binding protein (MBP) was produced with peptide15 fused to the N-terminus of MBP. The peptide15-MBP was expressed in E. coli and the purified fusion-protein was used to verify Kly18 specific binding. Chemically synthesised peptide15 (SWFPLRSGGG) could inhibit the enzymatic activity of both Kly18 and intact Karilysin (Kly48). Furthermore, peptide15 could slow down the autoprocessing of intact Kly48 to Kly18. The WFP motif was important for inhibition and a truncation study further demonstrated that the N-terminal serine was also essential for Kly18 inhibition. The SWFP peptide had a Ki value in the low micromolar range, which was similar to the intact peptide15. In conclusion SWFP is the first reported inhibitor of Karilysin and can be used as a valuable tool in structure-function studies of Karilysin.
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影响因子：
3.7
作者：
Ishihara K;Wawrzonek K;Shaw LN;Inagaki S;Miyamoto M;Potempa J
通讯作者：
Potempa J
影响因子：
4.8
作者：
Pan, WJ;Arnone, M;Albright, CF
通讯作者：
Albright, CF
影响因子：
2.2
作者：
RATH, S;STANLEY, CM;STEWARD, MW
通讯作者：
STEWARD, MW
DOI：
10.1093/protein/8.10.1063
发表时间：
1995-10-01
期刊：
PROTEIN ENGINEERING
影响因子：
--
作者：
Johansen, LK;Albrechtsen, B;Engberg, J
通讯作者：
Engberg, J
影响因子：
3.7
作者：
Krauser, JA;Potempa, J;Powers, JC
通讯作者：
Powers, JC