Author:
Xu Yueyang,Li Xin,Li Ruiqing,Li Shanshan,Ni Hongqian,Wang Hui,Xu Haijin,Zhou Weihong,Saris Per E. J.,Yang Wen,Qiao Mingqiang,Rao Zihe
Abstract
Nisin is a widely used antibacterial lantibiotic polypeptide produced byLactococcus lactis. NisP belongs to the subtilase family and functions in the last step of nisin maturation as the leader-peptide peptidase. Deletion of thenisPgene in LAC71 results in the production of a non-active precursor peptide with the leader peptide unremoved. Here, the 1.1 Å resolution crystal structure of NisP is reported. The structure shows similarity to other subtilases, which can bind varying numbers of Ca atoms. However, no calcium was found in this NisP structure, and the predicted calcium-chelating residues were placed so as to not allow NisP to bind a calcium ion in this conformation. Interestingly, a short peptide corresponding to its own 635–647 sequence was found to bind to the active site of NisP. Biochemical assays and native mass-spectrometric analysis confirmed that NisP possesses an auto-cleavage site between residues Arg647 and Ser648. Further, it was shown that NisP mutated at the auto-cleavage site (R647P/S648P) had full catalytic activity for nisin leader-peptide cleavage, although the C-terminal region of NisP was no longer cleaved. Expressing this mutant inL. lactisLAC71 did not affect the production of nisin but did decrease the proliferation rate of the bacteria, suggesting the biological significance of the C-terminal auto-cleavage of NisP.
Publisher
International Union of Crystallography (IUCr)
Subject
General Medicine,Structural Biology
Cited by
25 articles.
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