Co-Catalytic Metallopeptidases as Pharmaceutical Targets

Understanding the reaction mechanism of co-catalytic metallopeptidases provides a starting point for the design and synthesis of new molecules that can be screened as potential pharmaceuticals. Many of the enzymes that contain co-catalytic metallo-active sites play important roles in cellular processes such as tissue repair, protein maturation, hormone level regulation, cell-cycle control and protein degradation. Therefore, these enzymes play central roles in several disease states including cancer, HIV, stroke, diabetes, bacterial infections, neurological processes, schizophrenia, seizure disorders, and amyotrophic lateral sclerosis. The mechanism of AAP, an aminopeptidase from Aeromonas proteolytica, is one of the best-characterized examples of a metallopeptidase containing a co-catalytic metallo-active site, although this enzyme is not a specific pharmaceutical target at this time. As a large majority of co-catalytic metallopeptidases contain active sites that are nearly identical to the one observed in AAP, the major steps of their catalytic mechanisms are likely to be very similar. With this in mind, it is possible to propose a general catalytic mechanism for the hydrolysis of amino acid substrates

Light of Life

The path through life is filled with light and darkness but light casts shadows and can darken one\u27s life. With this metaphor, I use light to symbolize the positive and shadows to represent the negative things that every one of us encounters. In my landscape paintings, the combination of light and darkness symbolizes my journey through life. There is a predominance of light in my paintings. Therefore, my intention is to portray the positive over the negative. In other words, I want to portray the light at the end of the tunnel

MOTIVATION AND TEACHERS’ JOB SATISFACTION FOR IMPROVED ACADEMIC PERFORMANCE OF STUDENTS IN SECONDARY SCHOOLS IN ANANBRA STATE

This study investigated the motivation of teachers and their job satisfaction as correlates of improved students' academic performance in Secondary Schools in Anambra State. The total population for the study comprised 5,761 public secondary school teachers in Anambra State. The sample for the study comprised 1,728 teachers which were composed using a stratified random sampling technique. Two research questions guided the study and two hypotheses were tested at a 0.05 level of significance. The study utilized a correlational research design. The instrument was duly validated by experts. The researcher developed an instrument titled "Motivation and Teacher Job Satisfaction and Students' Academic Performance" Questionnaire (MTJSSAP) was used to collecting data.  A reliability index of 0.88 for the questionnaire was obtained using Cronbach’s Alpha to measure the internal consistency. Out of the 1,728 copies of the questionnaire distributed, I,700 copies were duly completed and retrieved representing a 98.4% return rate, and these were used for data analysis. Data were analyzed using Pearson's Product Moment Correlation Coefficient. The findings revealed that there is a positive correlation between motivation and improved students' academic performance.  It was therefore recommended that since teachers are happy with their job if they have clear information, professional achievement, recognition, opportunity to participate in decision making, receive a living wage, and have good relationships with their boss that the state government, and secondary school administrators should make efforts to uphold these conditions so as to improve teachers job satisfaction and ultimately students’ academic performance

Characterization of the Catalytically Active Mn(II)-loaded \u3cem\u3eargE\u3c/em\u3e-encoded \u3cem\u3eN\u3c/em\u3e-acetyl-L-ornithine Deacetylase from \u3cem\u3eEscherichia coli\u3c/em\u3e

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 calorimetry of 0.30 μM for the first metal binding event and 5.3 μM for the second. An Arrhenius plot of the NAO hydrolysis for Mn(II)-loaded ArgE was linear from 15 to 55 °C, suggesting the rate-limiting step does not change as a function of temperature over this range. The activation energy, determined from the slope of this plot, was 50.3 kJ mol−1. Other thermodynamic parameters were ΔG ‡ = 58.1 kJ mol−1, ΔH ‡ = 47.7 kJ mol−1, and ΔS ‡ = –34.5 J mol−1 K−1. Similarly, plots of lnK m versus 1/T were linear, suggesting substrate binding is controlled by a single step. The natural product, [(2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl]leucine (bestatin), was found to be a competitive inhibitor of ArgE with a K i value of 67 μM. Electron paramagnetic resonance (EPR) data recorded for both [Mn(II)_(ArgE)] and [Mn(II)Mn(II)(ArgE)] indicate that the two Mn(II) ions form a dinuclear site. Moreover, the EPR spectrum of [Mn(II)Mn(II)(ArgE)] in the presence of bestatin indicates that bestatin binds to ArgE but does not form a µ-alkoxide bridge between the two metal ions

\u3cem\u3eargE\u3c/em\u3e-Encoded \u3cem\u3eN\u3c/em\u3e-Acetyl-l-Ornithine Deacetylase from \u3cem\u3eEscherichia coli\u3c/em\u3e Contains a Dinuclear Metalloactive Site

The catalytic and structural properties of the argE-encoded N-acetyl-l-ornithine deacetylase (ArgE) from Escherichia coli were investigated. On the basis of kinetic and ITC (isothermal titration calorimetry) data, Zn(II) binds to ArgE with Kd values that differ by ∼20 times. Moreover, ArgE exhibits ∼90% of its full catalytic activity upon addition of one metal ion. Therefore, ArgE behaves similarly to the aminopeptidase from Aeromonas proteolytica (AAP) in that one metal ion is the catalytic metal ion while the second likely plays a structural role. The N-acetyl-l-ornithine (NAO) deacetylase activity of ArgE showed a linear temperature dependence from 20 to 45 °C, indicating that the rate-limiting step does not change over this temperature range. The activation energy for NAO hydrolysis by ArgE was 25.6 kJ/mol when loaded with Zn(II) and 34.3 kJ/mol when loaded with Co(II). Electronic absorption and EPR (electron paramagnetic resonance) spectra of [Co·(ArgE)] and [CoCo(ArgE)] indicate that both divalent metal binding sites are five coordinate. In addition, EPR data show clear evidence of spin−spin coupling between the Co(II) ions in the active site but only after addition of a second equivalent of Co(II). Combination of these data provides the first physical evidence that the ArgE from E. coli contains a dinuclear Zn(II) active site, similar to AAP and the carboxypeptidase G2 from Pseudomonas sp. strain RS-16 (CPG2)

Identification of a Histidine Metal Ligand in the \u3cem\u3eargE\u3c/em\u3e-Encoded \u3cem\u3eN\u3c/em\u3e-Acetyl-L-Ornithine Deacetylase from \u3cem\u3eEscherichia coli\u3c/em\u3e

The H355A, H355K, H80A, and H80K mutant enzymes of the argE-encoded N-acetyl-L-ornithine deacetylase (ArgE) from Escherichia coli were prepared, however, only the H355A enzyme was found to be soluble. Kinetic analysis of the Co(II)-loaded H355A exhibited activity levels that were 380-fold less than Co(II)-loaded WT ArgE. Electronic absorption spectra of Co(II)-loaded H355A-ArgE indicate that the bound Co(II) ion resides in a distorted, five-coordinate environment and Isothermal Titration Calorimetry (ITC) data for Zn(II) binding to the H355A enzyme provided a dissociation constant (Kd) of 39 μM. A three-dimensional homology model of ArgE was generated using the X-ray crystal structure of the dapE-encoded N-succinyl-L,L-diaminopimelic acid desuccinylase (DapE) from Haemophilus influenzae confirming the assignment of H355 as well as H80 as active site ligands

Galaxies as Fluctuations in the Ionizing Background Radiation at Low Redshift

Some Lyman continuum photons are likely to escape from most galaxies, and these can play an important role in ionizing gas around and between galaxies, including gas that gives rise to Lyman alpha absorption. Thus the gas surrounding galaxies and in the intergalactic medium will be exposed to varying amounts of ionizing radiation depending upon the distances, orientations, and luminosities of any nearby galaxies. The ionizing background can be recalculated at any point within a simulation by adding the flux from the galaxies to a uniform quasar contribution. Normal galaxies are found to almost always make some contribution to the ionizing background radiation at redshift zero, as seen by absorbers and at random points in space. Assuming that about 2 percent of ionizing photons escape from a galaxy like the Milky Way, we find that normal galaxies make a contribution of at least 30 to 40 percent of the assumed quasar background. Lyman alpha absorbers with a wide range of neutral column densities are found to be exposed to a wide range of ionization rates, although the distribution of photoionization rates for absorbers is found to be strongly peaked. On average, less highly ionized absorbers are found to arise farther from luminous galaxies, while local fluctuations in the ionization rate are seen around galaxies having a wide range of properties.Comment: 10 pages, 8 figures, references added, clarified explanation of first two equation