Molecular Genetics Dept., University of Groningen, Nijenborgh 7, 9747 AG, Groningen, The Netherlands ; Synthetic Biology Centre, University of Groningen, 9747 AG, Groningen, The Netherlands.
FEBS Open Bio. 2013 May 30;3:237-42. doi: 10.1016/j.fob.2013.05.001. Print 2013.
Nisin is the most prominent and applied bacteriocin that serves as a model for class I lantibiotics. The nisin leader peptide importantly determines interactions between precursor nisin and its modification enzymes NisB and NisC that mature nisin posttranslationally. NisB dehydrates serines and threonines, while NisC catalyzes the subsequent coupling of the formed dehydroamino acids to form lanthionines. Currently, little is known about how the nisin leader interacts with NisB and even less is known about its interactions with NisC. To investigate the nisin leader peptide requirements for functional interaction with the modification enzymes NisB and NisC, we systematically replaced six regions, of 2-4 amino acids each, with all-alanine regions. By performing NisB and NisC co-purification studies with these mutant leader peptides, we demonstrate that the nisin leader regions STKD(-22-19), FNLD(-18-15) and PR(-2-1) importantly contribute to the interactions of precursor nisin with both NisB and NisC, whereas the nisin leader region LVSV(-14-11) additionally contributes to the interaction of precursor nisin with NisC.
乳链菌肽是最显著和应用最广泛的细菌素，可用作 I 类羊毛硫抗生素的模型。乳链菌肽前导肽对于前体乳链菌肽与其修饰酶 NisB 和 NisC 的相互作用非常重要，后者在后翻译水平上成熟乳链菌肽。NisB 使丝氨酸和苏氨酸脱水，而 NisC 则催化形成的去氢氨基酸随后与形成的类硫氨酸结合。目前，人们对乳链菌肽前导肽与修饰酶 NisB 的相互作用知之甚少，甚至对其与 NisC 的相互作用也知之甚少。为了研究乳链菌肽前导肽与修饰酶 NisB 和 NisC 功能相互作用的要求，我们系统地用全丙氨酸区域替换了每个由 2-4 个氨基酸组成的六个区域。通过用这些突变的前导肽进行 NisB 和 NisC 共纯化研究，我们证明乳链菌肽前导肽的 STKD(-22-19)、FNLD(-18-15)和 PR(-2-1)区域对前体乳链菌肽与 NisB 和 NisC 的相互作用非常重要，而乳链菌肽前导肽 LVSV(-14-11)区域则对前体乳链菌肽与 NisC 的相互作用也很重要。
