PDBsum entry 1l9c

Oxidoreductase

PDB id: 1l9c

Contents

Protein chains

385 a.a. *

Ligands

FAD ×2

PO4

Metals

_CL ×2

Waters ×1065

* Residue conservation analysis

PDB id:

1l9c

Name:

Oxidoreductase

Title:

Role of histidine 269 in catalysis by monomeric sarcosine oxidase

Structure:

Monomeric sarcosine oxidase. Chain: a, b. Synonym: msox. Engineered: yes. Mutation: yes

Source:

Bacillus sp.. Organism_taxid: 69000. Strain: b-0618. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

1.90Å R-factor: 0.171 R-free: 0.216

Authors:

G.Zhao,H.Song,Z.-W.Chen,F.S.Mathews,M.S.Jorns

Key ref:

G.Zhao et al. (2002). Monomeric sarcosine oxidase: role of histidine 269 in catalysis. Biochemistry, 41, 9751-9764. PubMed id: 12146941 DOI: 10.1021/bi020286f

Date:

22-Mar-02 Release date: 30-Aug-02

Protein chains

P40859  (MSOX_BACB0) -  Monomeric sarcosine oxidase from Bacillus sp. (strain B-0618)

Seq: 390 a.a.

Struc: 385 a.a.*

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

Enzyme reactions

• Enzyme class: E.C.1.5.3.1 - sarcosine oxidasee (formaldehyde-forming).
• Reaction: sarcosine + O2 + H2O = formaldehyde + glycine + H2O2
• Cofactor: FAD

FAD (Bound ligand (Het Group name = FAD) corresponds exactly)

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

reference

DOI no: 10.1021/bi020286f

Biochemistry 41:9751-9764 (2002)

PubMed id: 12146941

Monomeric sarcosine oxidase: role of histidine 269 in catalysis.

G.Zhao, H.Song, Z.W.Chen, F.S.Mathews, M.S.Jorns.

ABSTRACT

Conservative mutation of His269 (to Asn, Ala, or Gln) does not-significantly affect the expression of monomeric sarcosine oxidase (MSOX), covalent flavinylation, the physicochemical properties of bound FAD, or the overall protein structure. Turnover with sarcosine and the limiting rate of the reductive half-reaction with L-proline at pH 8.0 are, however, nearly 2 orders of magnitude slower than that with with wild-type MSOX. The crystal structure of the His269Asn complex with pyrrole-2-carboxylate shows that the pyrrole ring of the inhibitor is displaced as compared with wild-type MSOX. The His269 mutants all form charge-transfer complexes with pyrrole-2-carboxylate or methylthioacetate, but the charge-transfer bands are shifted to shorter wavelengths (higher energy) as compared with wild-type MSOX. Both wild-type MSOX and the His269Asn mutant bind the zwitterionic form of L-proline. The E(ox).L-proline complex formed with the His269Asn mutant or wild-type MSOX contains an ionizable group (pK(a) = 8.0) that is required for conversion of the zwitterionic L-proline to the reactive anionic form, indicating that His269 is not the active-site base. We propose that the change in ligand orientation observed upon mutation of His269 results in a less than optimal overlap of the highest occupied orbital of the ligand with the lowest unoccupied orbital of the flavin. The postulated effect on orbital overlap may account for the increased energy of charge-transfer bands and the slower rates of electron transfer observed for mutant enzyme complexes with charge-transfer ligands and substrates, respectively.