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Articles 1 - 8 of 8
Full-Text Articles in Physics
Evidence For A Dinuclear Active Site In The Metallo-Β-Lactamase Bcii With Substoichiometric Co(Ii): A New Model For Uptake, Leticia L. Llarrull, Mariana F. Tioni, Jason M. Kowalski, Brian Bennett, Alejandro J. Vila
Evidence For A Dinuclear Active Site In The Metallo-Β-Lactamase Bcii With Substoichiometric Co(Ii): A New Model For Uptake, Leticia L. Llarrull, Mariana F. Tioni, Jason M. Kowalski, Brian Bennett, Alejandro J. Vila
Physics Faculty Research and Publications
Metallo-β-lactamases are zinc-dependent enzymes that constitute one of the main resistance mechanisms to β-lactam antibiotics. Metallo-β-lactamases have been characterized both in mono- and dimetallic forms. Despite many studies, the role of each metal binding site in substrate binding and catalysis is still unclear. This is mostly due to the difficulties in assessing the metal content and site occupancy in solution. For this reason, Co(II) has been utilized as a useful probe of the active site structure. We have employed UV-visible, EPR, and NMR spectroscopy to study Co(II) binding to the metallo-β-lactamase BcII from Bacillus cereus. The spectroscopic features were …
X-Ray Crystallographic Characterization Of The Co(Ii)-Substituted Tris-Bound Form Of The Aminopeptidase From Aeromonas Proteolytica, Petra Munih, Aaron Moulin, Carin Stamper, Brian Bennett, Dagmar Ringe, Gregory A. Petsko, Richard C. Holz
X-Ray Crystallographic Characterization Of The Co(Ii)-Substituted Tris-Bound Form Of The Aminopeptidase From Aeromonas Proteolytica, Petra Munih, Aaron Moulin, Carin Stamper, Brian Bennett, Dagmar Ringe, Gregory A. Petsko, Richard C. Holz
Physics Faculty Research and Publications
The X-ray crystal structure of the Co(II)-loaded form of the aminopeptidase from Aeromonas proteolytica ([CoCo(AAP)]) was solved to 2.2 Å resolution. [CoCo(AAP)] folds into an α/β globular domain with a twisted β-sheet hydrophobic core sandwiched between α-helices, identical to [ZnZn(AAP)]. Co(II) binding to AAP does not introduce any major conformational changes to the overall protein structure and the amino acid residues ligated to the dicobalt(II) cluster in [CoCo(AAP)] are the same as those in the native Zn(II)-loaded structure with only minor perturbations in bond lengths. The Co(II)–Co(II) distance is 3.3 Å. Tris(hydroxymethyl)aminomethane (Tris) coordinates to the dinuclear Co(II) active site …
Modified Active Site Coordination In A Clinical Mutant Of Sulfite Oxidase, Christian J. Doonan, Heather L. Wilson, K. V. Rajagopalan, Robert M. Garrett, Brian Bennett, Roger C. Prince, Graham N. George
Modified Active Site Coordination In A Clinical Mutant Of Sulfite Oxidase, Christian J. Doonan, Heather L. Wilson, K. V. Rajagopalan, Robert M. Garrett, Brian Bennett, Roger C. Prince, Graham N. George
Physics Faculty Research and Publications
The molybdenum site of the Arginine 160 → Glutamine clinical mutant of the physiologically vital enzyme sulfite oxidase has been investigated by a combination of X-ray absorption spectroscopy and density functional theory calculations. We conclude that the mutant enzyme has a six-coordinate pseudo-octahedral active site with coordination of Glutamine Oε to molybdenum. This contrasts with the wild-type enzyme which is five-coordinate with approximately square-based pyramidal geometry. This difference in the structure of the molybdenum site explains many of the properties of the mutant enzyme which have previously been reported.
The Metallo-Β-Lactamase Gob Is A Mono-Zn(Ii) Enzyme With A Novel Active Site, Jorgelina Morrán-Barrio, Javier M. Gonzalez, Mariá Natalia Lisa, Alison L. Costello, Matteo Dal Peraro, Paolo Carloni, Brian Bennett, David L. Tierney, Adriana S. Limansky, Alejandro M. Viale, Alejandro J. Vila
The Metallo-Β-Lactamase Gob Is A Mono-Zn(Ii) Enzyme With A Novel Active Site, Jorgelina Morrán-Barrio, Javier M. Gonzalez, Mariá Natalia Lisa, Alison L. Costello, Matteo Dal Peraro, Paolo Carloni, Brian Bennett, David L. Tierney, Adriana S. Limansky, Alejandro M. Viale, Alejandro J. Vila
Physics Faculty Research and Publications
Metallo-β-lactamases (MβLs) are zinc-dependent enzymes able to hydrolyze and inactivate most β-lactam antibiotics. The large diversity of active site structures and metal content among MβLs from different sources has limited the design of a pan-MβL inhibitor. Here we report the biochemical and biophysical characterization of a novel MβL, GOB-18, from a clinical isolate of a Gram-negative opportunistic pathogen, Elizabethkingia meningoseptica. Different spectroscopic techniques, three-dimensional modeling, and mutagenesis experiments, reveal that the Zn(II) ion is bound to Asp120, His121, His263, and a solvent molecule, i.e. in the canonical Zn2 site of dinuclear MβLs. Contrasting …
Asp-120 Locates Zn2 For Optimal Metallo-Β-Lactamase Activity, Leticia L. Llarrull, Stella M. Fabiane, Jason M. Kowalski, Brian Bennett, Brian J. Sutton, Alejandro J. Vila
Asp-120 Locates Zn2 For Optimal Metallo-Β-Lactamase Activity, Leticia L. Llarrull, Stella M. Fabiane, Jason M. Kowalski, Brian Bennett, Brian J. Sutton, Alejandro J. Vila
Physics Faculty Research and Publications
Metallo-β-lactamases are zinc-dependent hydrolases that inactivate β-lactam antibiotics, rendering bacteria resistant to them. Asp-120 is fully conserved in all metallo-β-lactamases and is central to catalysis. Several roles have been proposed for Asp-120, but so far there is no agreed consensus. We generated four site-specifically substituted variants of the enzyme BcII from Bacillus cereus as follows: D120N, D120E, D120Q, and D120S. Replacement of Asp-120 by other residues with very different metal ligating capabilities severely impairs the lactamase activity without abolishing metal binding to the mutated site. A kinetic study of these mutants indicates that Asp-120 is not the proton donor, nor …
Characterization Of The Catalytically Active Mn(Ii)-Loaded Arge-Encoded N-Acetyl-L-Ornithine Deacetylase From Escherichia Coli, Wade C. Mcgregor, Sabina I. Swierczek, Brian Bennett, Richard C. Holz
Characterization Of The Catalytically Active Mn(Ii)-Loaded Arge-Encoded N-Acetyl-L-Ornithine Deacetylase From Escherichia Coli, Wade C. Mcgregor, Sabina I. Swierczek, Brian Bennett, Richard C. Holz
Physics Faculty Research and Publications
The catalytically competent Mn(II)-loaded form of the argE-encoded N-acetyl-l-ornithine deacetylase from Escherichia coli (ArgE) was characterized by kinetic, thermodynamic, and spectroscopic methods. Maximum N-acetyl-l-ornithine (NAO) hydrolytic activity was observed in the presence of one Mn(II) ion with k cat and K m values of 550 s−1 and 0.8 mM, respectively, providing a catalytic efficiency (k cat/K m) of 6.9 × 105 M−1 s−1. The ArgE dissociation constant (K d) for Mn(II) was determined to be 0.18 μM, correlating well with a value obtained by isothermal titration …
Micromagnetics Of The Domain Wall Mobility In Permalloy Nanowires, Andrew Kunz
Micromagnetics Of The Domain Wall Mobility In Permalloy Nanowires, Andrew Kunz
Physics Faculty Research and Publications
The domain wall mobility in long permalloy nanowires with thicknesses of 2-20 nm and widths of 50-200 nm has been simulated. The domain wall is driven into motion by an external magnetic field and the average wall mobility is calculated after the wall has traveled 2.5 mum along the wire. The results were obtained using the three-dimensional dynamic Landau-Lifshitz equation. We find that the domain wall mobility decreases linearly up to the critical field called the Walker field. The decreasing wall mobility is related to the decrease in the dynamic domain wall length as the applied field is increased. The …
Experimental Evidence For A Metallohydrolase Mechanism In Which The Nucleophile Is Not Delivered By A Metal Ion: Epr Spectrokinetic And Structural Studies Of Aminopeptidase From Vibrio Proteolyticus, Amit Kumar, Gopal R. Periyannan, Aaron W. Kittell, Jung Ja Kim, Brian Bennett
Experimental Evidence For A Metallohydrolase Mechanism In Which The Nucleophile Is Not Delivered By A Metal Ion: Epr Spectrokinetic And Structural Studies Of Aminopeptidase From Vibrio Proteolyticus, Amit Kumar, Gopal R. Periyannan, Aaron W. Kittell, Jung Ja Kim, Brian Bennett
Physics Faculty Research and Publications
Metallohydrolases catalyse some of the most important reactions in biology and are targets for numerous chemotherapeutic agents designed to combat bacterial infectivity, antibiotic resistance, HIV infectivity, tumour growth, angiogenesis and immune disorders. Rational design of inhibitors of these enzymes with chemotherapeutic potential relies on detailed knowledge of the catalytic mechanism. The roles of the catalytic transition ions in these enzymes have long been assumed to include the activation and delivery of a nucleophilic hydroxy moiety. In the present study, catalytic intermediates in the hydrolysis of L-leucyl-L-leucyl-L-leucine by Vibrio proteolyticus aminopeptidase were characterized in spectrokinetic and structural studies. Rapid-freeze-quench EPR studies …