Satomura Atsushi, Kuroda Kouichi, Ueda Mitsuyoshi
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, Japan; Japan Society for the Promotion of Science, Sakyo-ku, Kyoto, Japan.
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, Japan.
PLoS One. 2015 May 13;10(5):e0124545. doi: 10.1371/journal.pone.0124545. eCollection 2015.
Rhizopus oryzae lipase (ROL) has a propeptide at its N-terminus that functions as an intramolecular chaperone and facilitates the folding of mature ROL (mROL). In this study, we successfully generated a functionally distinct imprinted mROL (mROLimp) through protein folding memory using a mutated propeptide. The mutated propeptide left its structural memory on mROL and produced mROLimp that exhibited different substrate specificities compared with mROLWT (prepared from the wild type propeptide), although the amino acid sequences of both mROLs were the same. mROLimp showed a preference for substrates with medium chain-length acyl groups and, noticeably, recognized a peptidase-specific substrate. In addition, ROLimp was more stable than mROLWT. These results strongly suggest that proteins with identical amino acid sequences can fold into different conformations and that mutations in intramolecular chaperones can dynamically induce changes in enzymatic activity.
米根霉脂肪酶（ROL）在其N端有一个前肽，该前肽作为分子内伴侣发挥作用，促进成熟ROL（mROL）的折叠。在本研究中，我们通过使用突变前肽的蛋白质折叠记忆成功产生了功能不同的印记mROL（mROLimp）。突变前肽在mROL上留下了其结构记忆，并产生了与mROLWT（由野生型前肽制备）相比表现出不同底物特异性的mROLimp，尽管两种mROL的氨基酸序列相同。mROLimp对具有中链长度酰基的底物表现出偏好，并且值得注意的是，它识别一种肽酶特异性底物。此外，ROLimp比mROLWT更稳定。这些结果有力地表明，具有相同氨基酸序列的蛋白质可以折叠成不同的构象，并且分子内伴侣中的突变可以动态地诱导酶活性的变化。
