Effects of Phosphorylation on Phosphoenolpyruvate Carboxykinase from the C4 Plant Guinea Grass

In the C4 plant Guinea grass (Panicum maximum), phosphoenolpyruvate carboxykinase (PEPCK) is phosphorylated in darkened leaves and dephosphorylated in illuminated leaves. To determine whether the properties of phosphorylated and non-phosphorylated PEPCK were different, PEPCK was purified to homogeneity from both illuminated and darkened leaves. The final step of the purification procedure, gel filtration chromatography, further separated phosphorylated and non-phosphorylated forms. In the presence of a high ratio of ATP to ADP, the non-phosphorylated enzyme had a higher affinity for its substrates, oxaloacetate and phosphoenolpyruvate. The activity of the non-phosphorylated form was up to 6-fold higher when measured at low substrate concentrations. Comparison of proteoloytically cleaved PEPCK from Guinea grass, which lacked its N-terminal extension, from yeast (Saccharomyces cerevisiae), which does not possess an N-terminal extension, and from the C4 plant Urochloa panicoides, which possesses an N-terminal extension but is not subject to phosphorylation, revealed similar properties to the non-phosphorylated full-length form from Guinea grass. Assay of PEPCK activity in crude extracts of Guinea grass leaves, showed a large difference between illuminated and darkened leaves when measured in a selective assay (a low concentration of phosphoenolpyruvate and a high ratio of ATP to ADP), but there was no difference under assay conditions used to estimate maximum activity. Immunoblots of sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels showed no difference in the abundance of PEPCK protein in illuminated and darkened leaves. There were no light/dark differences in activity detected in maize (Zea mays) leaves, in which PEPCK is not subject to phosphorylation

Phosphoenolpyruvate Carboxykinase Assayed at Physiological Concentrations of Metal Ions Has a High Affinity for CO2

The effect of Mn2+/Mg2+ concentration on the activity of intact, homogeneous phosphoenolpyruvate carboxykinase (PEPCK) from leaves of the C4 grass, Guinea grass (Panicum maximum), have been investigated. Assay conditions were optimized so that PEPCK activity could be measured at concentrations of Mn2+/Mg2+ similar to those found in the cytosol (low micromolar Mn2+ and millimolar Mg2+). PEPCK activity was totally dependent on Mn2+ and was activated at low micromolar concentrations of Mn2+ by millimolar concentrations of Mg2+. Therefore, at physiological concentrations of Mn2+, PEPCK has a requirement for Mg2+. Assay at physiological concentrations of Mn2+/Mg2+ led to a marked decrease in its affinity for ATP and a 13-fold increase in its affinity for CO2. The Km (CO2) was further decreased by assay at physiological ATP to ADP ratios, reaching values as low as 20 μM CO2, comparable with the Km (CO2) of ribulose 1,5-bisphosphate carboxylase-oxygenase. This means that PEPCK will catalyze a reversible reaction and that it could operate as a carboxylase in vivo, a feature that could be particularly important in algal CO2-concentrating systems

Project pathogens: The anatomy of omission errors in construction and resource engineering project

Construction and engineering projects are typically complex in nature and are prone to cost and schedule overruns. A significant factor that often contributes to these overruns is rework. Omissions errors, in particular, have been found to account for as much as 38% of the total rework costs experienced. To date, there has been limited research that has sought to determine the underlying factors that contribute to omission errors in construction and engineering projects. Using data derived from59 in-depth interviews undertaken with various project participants, a generic systemic causal model of the key factors that contributed to omission errors is presented. The developed causal model can improve understanding of the archetypal nature and underlying dynamics of omission errors. Error management strategies that can be considered for implementation in projects are also discussed

Phosphoenolpyruvate Carboxykinase Is Involved in the Decarboxylation of Aspartate in the Bundle Sheath of Maize

We recently showed that maize (Zea mays L.) leaves contain appreciable amounts of phosphoenolpyruvate carboxykinase (PEPCK; R.P. Walker, R.M. Acheson, L.I. Técsi, R.C. Leegood [1997] Aust J Plant Physiol 24: 459–468). In the present study, we investigated the role of PEPCK in C4 photosynthesis in maize. PEPCK activity and protein were enriched in extracts from bundle-sheath (BS) strands compared with whole-leaf extracts. Decarboxylation of [4-14C]aspartate (Asp) by BS strands was dependent on the presence of 2-oxoglutarate and Mn2+, was stimulated by ATP, was inhibited by the PEPCK-specific inhibitor 3-mercaptopicolinic acid, and was independent of illumination. The principal product of Asp metabolism was phosphoenolpyruvate, whereas pyruvate was a minor product. Decarboxylation of [4-14C]malate was stimulated severalfold by Asp and 3-phosphoglycerate, was only slightly reduced in the absence of Mn2+ or in the presence of 3-mercaptopicolinic acid, and was light dependent. Our data show that decarboxylation of Asp and malate in BS cells of maize occurs via two different pathways: Whereas malate is mainly decarboxylated by NADP-malic enzyme, decarboxylation of Asp is dependent on the activity of PEPCK

Community College Culture and Faculty of Color

This investigation examines and explains the ways in which community college faculty of color construct their understandings of institutional culture. We investigate four community colleges in California through interviews with 31 full-time faculty of color. This faculty group expresses identity conflicts between their professional roles and their cultural identities. Their understandings of their institutions suggest that the culture of the community college is more complex and multi-faceted than that portrayed in the scholarly literature, which often portrays the institution as homogeneous and the faculty body as uniform. © The Author(s) 2013

The Divided Self: The Double Consciousness of Faculty of Color in Community Colleges

Through qualitative field methods research addressing faculty of color in four California community colleges, this investigation examines and explains faculty experiences and professional sense making. By combining critical race theory with social identity theory, our perspective underlines the potential social and ethnic identity conflicts inherent in the daily lives of faculty of color. The professional and social identities of faculty of color are not necessarily compatible, leading to a condition of "double consciousness," or what we refer to as "the divided self." © The Author(s) 2013

“Dangerous Work”: Improving Conditions for Faculty of Color in the Community College

This qualitative investigation of the experiences of faculty of color at community colleges identifies current conditions for this population and suggests potentials for ameliorating conditions that inhibit their job satisfaction. We argue that the current conditions for faculty of color, based upon their expressed experiences at the community colleges, are deleterious to their professional performance, to their positive self-image, and to their contributions to their institutions. Alterations to these current conditions are unlikely without systemic institutional change. Indeed, without improvement to these conditions, the job satisfaction of faculty of color is not likely to change

An Equitable Educational Experience For All: Strategies For Secondary General Education Teachers to Meet the Learning Needs of Special Education Students Learning in Their Classrooms

This capstone project focuses on the issue of providing equitable educational experiences for all students. More specifically, the project examines the most effective strategies for secondary general education teachers to provide modifications, accommodations, and differentiation for special education students in the areas of instruction and assessment. A central tenet of contemporary special education policy is the principle of inclusion. In accordance with this principle, students with special needs are increasingly learning in the general education classroom among their peers. This places an additional responsibility upon general education teachers to ensure that they are meeting the unique learning needs of special education students learning in their classrooms. This capstone project seeks to equip general education teachers with a useful strategy for providing modifications, accommodations, and differentiation in the instruction and assessment that they deliver to their students with special needs. The project is designed as a professional development workshop that consists of four different sessions taking place throughout the first semester of the school year. During the initial session, secondary general education teachers are introduced to the I.N.C.L.U.D.E. strategy and have the opportunity to practice applying it to a hypothetical classroom situation. Over the course of the following three sessions, teachers reflect upon their own use of the strategy throughout the first semester. The utility and effectiveness of the I.N.C.L.U.D.E. strategy is measured by comparing teacher responses to a survey administered prior to the first session with teacher responses to the same survey after the final session

Report on Phase Two: The readiness to practise of NQSWs.

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