PDBsum entry 4mzd

Hydrolase

PDB id: 4mzd

Contents

Protein chain

356 a.a.

Waters ×454

PDB id:

4mzd

Name:

Hydrolase

Title:

High resolution crystal structure of the nisin leader peptidase nisp from lactococcus lactis

Structure:

Nisin leader peptide-processing serine protease nisp. Chain: a. Fragment: unp residues 196-682. Engineered: yes

Source:

Lactococcus lactis subsp. Lactis. Organism_taxid: 1360. Gene: nisp. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

1.10Å R-factor: 0.153 R-free: 0.168

Authors:

Z.H.Rao,Y.Y.Xu,X.Li,W.Yang

Key ref:

Y.Xu et al. (2014). Structure of the nisin leader peptidase NisP revealing a C-terminal autocleavage activity. Acta Crystallogr D Biol Crystallogr, 70, 1499-1505. PubMed id: 24914961 DOI: 10.1107/S1399004714004234

Date:

30-Sep-13 Release date: 11-Jun-14

Protein chain

Q07596  (NISP_LACLL) -  Nisin leader peptide-processing serine protease NisP from Lactococcus lactis subsp. lactis

Seq: 682 a.a.

Struc: 356 a.a.*

* PDB and UniProt seqs differ at 13 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.3.4.21.- - ?????

DOI no: 10.1107/S1399004714004234

Acta Crystallogr D Biol Crystallogr 70:1499-1505 (2014)

PubMed id: 24914961

Structure of the nisin leader peptidase NisP revealing a C-terminal autocleavage activity.

Y.Xu, X.Li, R.Li, S.Li, H.Ni, H.Wang, H.Xu, W.Zhou, P.E.Saris, W.Yang, M.Qiao, Z.Rao.

ABSTRACT

Nisin is a widely used antibacterial lantibiotic polypeptide produced by Lactococcus lactis. NisP belongs to the subtilase family and functions in the last step of nisin maturation as the leader-peptide peptidase. Deletion of the nisP gene 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 in L. lactis LAC71 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.