title: Inhibition of the dapE-Encoded N-Succinyl-L,L-diaminopimelic Acid Desuccinylase from Neisseria meningitidis by L-Captopril.
abstract: Binding of the competitive inhibitor L-captopril to the dapE-encoded N-succinyl-L,L-diaminopimelic acid desuccinylase from Neisseria meningitidis (NmDapE) was examined by kinetic, spectroscopic, and crystallographic methods. L-Captopril, an angiotensin-converting enzyme (ACE) inhibitor, was previously shown to be a potent inhibitor of the DapE from Haemophilus influenzae (HiDapE) with an IC50 of 3.3 μM and a measured Ki of 1.8 μM and displayed a dose-responsive antibiotic activity toward Escherichia coli. L-Captopril is also a competitive inhibitor of NmDapE with a Ki of 2.8 μM. To examine the nature of the interaction of L-captopril with the dinuclear active site of DapE, we have obtained electron paramagnetic resonance (EPR) and magnetic circular dichroism (MCD) data for the enzymatically hyperactive Co(II)-substituted forms of both HiDapE and NmDapE. EPR and MCD data indicate that the two Co(II) ions in DapE are antiferromagnetically coupled, yielding an S = 0 ground state, and suggest a thiolate bridge between the two metal ions. Verification of a thiolate-bridged dinuclear complex was obtained by determining the three-dimensional X-ray crystal structure of NmDapE in complex with L-captopril at 1.8 Å resolution. Combination of these data provides new insights into binding of L-captopril to the active site of DapE enzymes as well as important inhibitor-active site residue interaction's. Such information is critical for the design of new, potent inhibitors of DapE enzymes.
authors.affiliation: †Department of Chemistry and Biochemistry, Loyola University-Chicago, 1068 West Sheridan Road, Chicago, Illinois 60626, United States.
authors.forename: Anna
authors.initials: A
authors.lastname: Starus
authors.affiliation: ‡Center for Structural Genomics of Infectious Diseases, Computation Institute, University of Chicago, 5735 South Ellis Avenue, Chicago, Illinois 60637, United States.
authors.forename: Boguslaw
authors.initials: B
authors.lastname: Nocek
authors.affiliation: §Department of Physics, Marquette University, Milwaukee, Wisconsin 53201, United States.
authors.forename: Brian
authors.initials: B
authors.lastname: Bennett
authors.affiliation: ∥Department of Chemistry and Biochemistry, Middlebury College, Middlebury, Vermont 05753, United States.
authors.forename: James A
authors.initials: JA
authors.lastname: Larrabee
authors.affiliation: ∥Department of Chemistry and Biochemistry, Middlebury College, Middlebury, Vermont 05753, United States.
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Shaw
authors.affiliation: ∥Department of Chemistry and Biochemistry, Middlebury College, Middlebury, Vermont 05753, United States.
authors.forename: Wisath
authors.initials: W
authors.lastname: Sae-Lee
authors.affiliation: ∥Department of Chemistry and Biochemistry, Middlebury College, Middlebury, Vermont 05753, United States.
authors.forename: Marie T
authors.initials: MT
authors.lastname: Russo
authors.affiliation: ⊥Department of Chemistry, Silesian University of Technology, 44-100 Gliwice, Poland.
authors.forename: Danuta M
authors.initials: DM
authors.lastname: Gillner
authors.affiliation: ‡Center for Structural Genomics of Infectious Diseases, Computation Institute, University of Chicago, 5735 South Ellis Avenue, Chicago, Illinois 60637, United States.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Makowska-Grzyska
authors.affiliation: ‡Center for Structural Genomics of Infectious Diseases, Computation Institute, University of Chicago, 5735 South Ellis Avenue, Chicago, Illinois 60637, United States.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Joachimiak
authors.affiliation: #Department of Chemistry, Marquette University, Milwaukee, Wisconsin 53201, United States.
authors.forename: Richard C
authors.initials: RC
authors.lastname: Holz
mesh_terms: D000806:Angiotensin-Converting Enzyme Inhibitors / Q000737:chemistry*; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry; D002216:Captopril / Q000737:chemistry*; D020134:Catalytic Domain; D002942:Circular Dichroism; D018360:Crystallography, X-Ray; D008190:Lyases / Q000037:antagonists & inhibitors / Q000737:chemistry; D009345:Neisseria meningitidis / Q000201:enzymology*
chemical_list: D000806:Angiotensin-Converting Enzyme Inhibitors; D001426:Bacterial Proteins; D002216:Captopril; D008190:Lyases