PDBsum entry 2nrs

Biosynthetic protein

PDB id: 2nrs

Contents

Protein chains

403 a.a. *

* Residue conservation analysis

PDB id:

2nrs

Name:

Biosynthetic protein

Title:

Moea s371w

Structure:

Molybdopterin biosynthesis protein moea. Chain: a, b. Engineered: yes. Mutation: yes

Source:

Escherichia coli. Organism_taxid: 562. Gene: moea, bisb, chle, nare. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.80Å R-factor: 0.250 R-free: 0.303

Authors:

J.Nicolas,S.Xiang,H.Schindelin,K.V.Rajagopalan

Key ref:

J.D.Nichols et al. (2007). Mutational analysis of Escherichia coli MoeA: two functional activities map to the active site cleft. Biochemistry, 46, 78-86. PubMed id: 17198377 DOI: 10.1021/bi061551q

Date:

02-Nov-06 Release date: 16-Jan-07

Protein chains

P12281  (MOEA_ECOLI) -  Molybdopterin molybdenumtransferase from Escherichia coli (strain K12)

Seq: 411 a.a.

Struc: 403 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.2.10.1.1 - molybdopterin molybdotransferase.
• Reaction: adenylyl-molybdopterin + molybdate = Mo-molybdopterin + AMP + H⁺
• Cofactor: Zn(2+) or Mg(2+)

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

Key reference

DOI no: 10.1021/bi061551q

Biochemistry 46:78-86 (2007)

PubMed id: 17198377

Mutational analysis of Escherichia coli MoeA: two functional activities map to the active site cleft.

J.D.Nichols, S.Xiang, H.Schindelin, K.V.Rajagopalan.

ABSTRACT

The molybdenum cofactor is ubiquitous in nature, and the pathway for Moco biosynthesis is conserved in all three domains of life. Recent work has helped to illuminate one of the most enigmatic steps in Moco biosynthesis, ligation of metal to molybdopterin (the organic component of the cofactor) to form the active cofactor. In Escherichia coli, the MoeA protein mediates ligation of Mo to molybdopterin while the MogA protein enhances this process in an ATP-dependent manner. The X-ray crystal structures for both proteins have been previously described as well as two essential MogA residues, Asp49 and Asp82. Here we describe a detailed mutational analysis of the MoeA protein. Variants of conserved residues at the putative active site of MoeA were analyzed for a loss of function in two different, previously described assays, one employing moeA- crude extracts and the other utilizing a defined system. Oddly, no correlation was observed between the activity in the two assays. In fact, our results showed a general trend toward an inverse relationship between the activity in each assay. Moco binding studies indicated a strong correlation between a variant's ability to bind Moco and its activity in the purified component assay. Crystal structures of the functionally characterized MoeA variants revealed no major structural changes, indicating that the functional differences observed are not due to disruption of the protein structure. On the basis of these results, two different functional areas were assigned to regions at or near the MoeA active site cleft.