Structural basis for catalysis by the mono- and dimetalated forms of the dapE-encoded N-succinyl-L,L-diaminopimelic acid desuccinylase.
Biosynthesis of lysine and meso-diaminopimelic acid in bacteria provides essential components for protein synthesis and construction of the bacterial peptidoglycan cell wall. The dapE operon enzymes synthesize both meso-diaminopimelic acid and lysine and, therefore, represent potential targets for novel antibacterials. The dapE-encoded N-succinyl-L,L-diaminopimelic acid desuccinylase functions in a late step of the pathway and converts N-succinyl-L,L-diaminopimelic acid to L,L-diaminopimelic acid and succinate. Deletion of the dapE gene is lethal to Helicobacter pylori and Mycobacterium smegmatis, indicating that DapE's are essential for cell growth and proliferation. Since there are no similar pathways in humans, inhibitors that target DapE may have selective toxicity against only bacteria. A major limitation in developing antimicrobial agents that target DapE has been the lack of structural information. Herein, we report the high-resolution X-ray crystal structures of the DapE from Haemophilus influenzae with one and two zinc ions bound in the active site, respectively. These two forms show different activity. Based on these newly determined structures, we propose a revised catalytic mechanism of peptide bond cleavage by DapE enzymes. These structures provide important insight into catalytic mechanism of DapE enzymes as well as a structural foundation that is critical for the rational design of DapE inhibitors.
Nocek
Boguslaw P
BP
Midwest Center for Structural Genomics and Structural Biology Center, Argonne National Laboratory, Argonne, IL 60439, USA.
Gillner
Danuta M
DM
Fan
Yao
Y
Holz
Richard C
RC
Joachimiak
Andrzej
A
D000581:Amidohydrolases / Q000737:chemistry* / Q000378:metabolism; D002384:Catalysis; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D003960:Diaminopimelic Acid / Q000737:chemistry* / Q000378:metabolism; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism; D006193:Haemophilus influenzae / Q000201:enzymology* / Q000254:growth & development; D008956:Models, Chemical; D011487:Protein Conformation; D015032:Zinc / Q000737:chemistry*
D000581:Amidohydrolases; D003960:Diaminopimelic Acid; D004791:Enzyme Inhibitors; D015032:Zinc; C054685:N-succinyl-l,l-diaminopimelic acid desuccinylase