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Article
The dapE-encoded N-succinyl-l,l-diaminopimelic Acid Desuccinylase from Haemophilus influenzae Contains Two Active-site Histidine Residues
Journal of Biological Inorganic Biochemistry
  • Danuta Gillner, Loyola University Chicago
  • David L. Bienvenue, Loyola University Chicago
  • Boguslaw P. Nocek, Argonne National Laboratory
  • Andzrej Joachimiak, Argonne National Laboratory
  • Vincentos Zachary, Loyola University Chicago
  • Brian Bennett, Marquette University
  • Richard C. Holz, Marquette University
Document Type
Article
Language
eng
Format of Original
10 p.
Publication Date
1-1-2009
Publisher
Springer
Original Item ID
DOI: 10.1007/s00775-008-0418-z, PubMed Central: PMC2678232
Disciplines
Abstract

The catalytic and structural properties of the H67A and H349A dapE-encoded N-succinyl-l,l-diaminopimelic acid desuccinylase (DapE) from Haemophilus influenzae were investigated. On the basis of sequence alignment with the carboxypeptidase from Pseudomonas sp. strain RS-16, both H67 and H349 were predicted to be Zn(II) ligands. The H67A DapE enzyme exhibited a decreased catalytic efficiency (180-fold) compared with wild-type (WT) DapE towards N-succinyldiaminopimelic acid. No catalytic activity was observed for H349A under the experimental conditions used. The electronic paramagnetic resonance (EPR) and electronic absorption data indicate that the Co(II) ion bound to H349A-DapE is analogous to that of WT DapE after the addition of a single Co(II) ion. The addition of 1 equiv of Co(II) to H67A DapE provides spectra that are very different from those of the first Co(II) binding site of the WT enzyme, but that are similar to those of the second binding site. The EPR and electronic absorption data, in conjunction with the kinetic data, are consistent with the assignment of H67 and H349 as active-site metal ligands for the DapE from H. influenzae. Furthermore, the data suggest that H67 is a ligand in the first metal binding site, while H349 resides in the second metal binding site. A three-dimensional homology structure of the DapE from H. influenzae was generated using the X-ray crystal structure of the DapE from Neisseria meningitidis as a template and superimposed on the structure of the aminopeptidase from Aeromonas proteolytica (AAP). This homology structure confirms the assignment of H67 and H349 as active-site ligands. The superimposition of the homology model of DapE with the dizinc(II) structure of AAP indicates that within 4.0 Å of the Zn(II) binding sites of AAP all of the amino acid residues of DapE are nearly identical.

Comments

Accepted version. Journal of Biological Inorganic Biochemistry, Vol.14, No. 1 (January 1, 2009): 1-10. DOI. © SBIC 2008. Used with permission.

“The final publication is available at Springer via http://dx.doi.org/10.1007/s00775-008-0418-z”.

Brian Bennett was affiliated with Medical College of Wisconsin at the time of publication.

Richard C. Holz was affiliated with Loyola University-Chicago at the time of publication.

Citation Information
Danuta Gillner, David L. Bienvenue, Boguslaw P. Nocek, Andzrej Joachimiak, et al.. "The dapE-encoded N-succinyl-l,l-diaminopimelic Acid Desuccinylase from Haemophilus influenzae Contains Two Active-site Histidine Residues" Journal of Biological Inorganic Biochemistry (2009) ISSN: 0949-8257
Available at: http://works.bepress.com/richard_holz/39/