Molecular characterization of a prolyl endopeptidase from a feather-degrading thermophile Meiothermus ruber H328
Molecular characterization of a prolyl endopeptidase from a feather-degrading thermophile Meiothermus ruber H328
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羽毛降解嗜热菌 Meiothermus ruber H328 脯氨酰内肽酶的分子特征
DOI：
10.1093/jb/mvaa069
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Kunihiko Watanabe
中科院分区：
文献类型：
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作者：
Fumi Yamamoto; Hironobu Morisaka; Mitsuyoshi Ueda; Kunihiko Watanabe
脯氨酰内肽酶从好氧和革兰氏阴性嗜热Meiothermus ruberH 328（MrPEP）纯化的天然和重组形式，但两种制剂具有可比的特性。通过添加完整的鸡毛，天然MrPEP的产量增加了10倍。从菌株H328的基因组中克隆了MrPEP的基因（mrH_2860），发现在N-末端没有信号序列。MrPEP由两个主要结构域组成：β-螺旋桨结构域和肽酶结构域，具有典型的活性位点基序和催化三联体。基于对不同类型的肽底物和FRETS-25 Xaa文库的广泛研究，MrPEP在底物特异性上显示出对P1位置的Pro残基的严格偏好，但对P2和P3位置的更广泛偏好，对长度大于4个残基的底物肽的P3位置的残基具有更强的亲和力。总之，MrPEP的分子表征在功能和结构上与其动物对应物比细菌对应物更接近。
Prolyl endopeptidase from an aerobic and Gram-negative thermophileMeiothermus ruberH328 (MrPEP) was purified in native and recombinant forms, but both preparations had comparable characteristics. Production of the native MrPEP was increased 10-fold by adding intact chicken feathers. The gene for MrPEP (mrH_2860) was cloned from the genome of strain H328 and found to have no signal sequence at the N-terminus. MrPEP is composed of two major domains: the β-propeller domain and the peptidase domain with a typical active site motif and catalytic triad. Based on extensive investigations with different types of peptide substrates and FRETS-25Xaa libraries, MrPEP showed strict preferences for Pro residue at the P1 position but broader preferences at the P2 and P3 positions in substrate specificity with stronger affinity for residues at the P3 position of substrate peptides that are longer than four residues in length. In conclusion, the molecular characterization of MrPEP resembles its animal counterparts more closely than bacterial counterparts in function and structure.