PDBsum entry 4gr5

Ligase

PDB id: 4gr5

Contents

Protein chains

451 a.a.

Ligands

APC ×4

TLA ×6

Metals

_CL

Waters ×1061

PDB id:

4gr5

Name:

Ligase

Title:

Crystal structure of slgn1deltaasub in complex with ampcpp

Structure:

Non-ribosomal peptide synthetase. Chain: a, b, c, d. Engineered: yes

Source:

Streptomyces lydicus. Organism_taxid: 47763. Gene: slgn1. Expressed in: escherichia coli. Expression_system_taxid: 469008.

Resolution:

1.92Å R-factor: 0.159 R-free: 0.186

Authors:

D.A.Herbst,G.Zocher,T.Stehle

Key ref:

D.A.Herbst et al. (2013). Structural basis of the interaction of MbtH-like proteins, putative regulators of nonribosomal peptide biosynthesis, with adenylating enzymes. J Biol Chem, 288, 1991-2003. PubMed id: 23192349

Date:

24-Aug-12 Release date: 28-Nov-12

Protein chains

D1GLU5  (D1GLU5_9ACTN) -  Non-ribosomal peptide synthetase from Streptomyces lydicus

Seq: 567 a.a.

Struc: 451 a.a.

Enzyme reactions

• Enzyme class: E.C.?

J Biol Chem 288:1991-2003 (2013)

PubMed id: 23192349

Structural basis of the interaction of MbtH-like proteins, putative regulators of nonribosomal peptide biosynthesis, with adenylating enzymes.

D.A.Herbst, B.Boll, G.Zocher, T.Stehle, L.Heide.

ABSTRACT

The biosynthesis of nonribosomally formed peptides (NRPs), which include important antibiotics such as vancomycin, requires the activation of amino acids through adenylate formation. The biosynthetic gene clusters of NRPs frequently contain genes for small, so-called MbtH-like proteins. Recently, it was discovered that these MbtH-like proteins are required for some of the adenylation reactions in NRP biosynthesis, but the mechanism of their interaction with the adenylating enzymes has remained unknown. In this study, we determined the structure of SlgN1, a 3-methylaspartate-adenylating enzyme involved in the biosynthesis of the hybrid polyketide/NRP antibiotic streptolydigin. SlgN1 contains an MbtH-like domain at its N terminus, and our analysis defines the parameters required for an interaction between MbtH-like domains and an adenylating enzyme. Highly conserved tryptophan residues of the MbtH-like domain critically contribute to this interaction. Trp-25 and Trp-35 form a cleft on the surface of the MbtH-like domain, which accommodates the alanine side chain of Ala-433 of the adenylating domain. Mutation of Ala-433 to glutamate abolished the activity of SlgN1. Mutation of Ser-23 of the MbtH-like domain to tyrosine resulted in strongly reduced activity. However, the activity of this S23Y mutant could be completely restored by addition of the intact MbtH-like protein CloY from another organism. This suggests that the interface found in the structure of SlgN1 is the genuine interface between MbtH-like proteins and adenylating enzymes.