Digital Commons Network^™

The Role Of Surface Chemistry-Induced Cell Characteristics On Nonviral Gene Delivery To Mouse Fibroblasts, Tadas Kasputis, Angela K. Pannier

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Background: Gene delivery approaches serve as a platform to modify gene expression of a cell population with applications including functional genomics, tissue engineering, and gene therapy. The delivery of exogenous genetic material via nonviral vectors has proven to be less toxic and to cause less of an immune response in comparison to viral vectors, but with decreased efficiency of gene transfer. Attempts have been made to improve nonviral gene transfer efficiency by modifying physicochemical properties of gene delivery vectors as well as developing new delivery techniques. In order to further improve and understand nonviral gene delivery, our approach focuses …

Fabrication And Characterization Of Dna-Loaded Zein Nanospheres, Mary C. Regier, Jessica D. Taylor, Tyler Borcyk, Yiqi Yang, Angela K. Pannier

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Background: Particulates incorporating DNA are promising vehicles for gene delivery, with the ability to protect DNA and provide for controlled, localized, and sustained release and transfection. Zein, a hydrophobic protein from corn, is biocompatible and has properties that make it a promising candidate material for particulate delivery, including its ability to form nanospheres through coacervation and its insolubility under physiological conditions, making it capable of sustained release of encapsulated compounds. Due to the promise of this natural biomaterial for drug delivery, the objective of this study was to formulate zein nanospheres encapsulating DNA as the therapeutic compound, and to …

Runoff Nutrient Loads As Affected By Residue Cover, Manure Application Rate, And Flow Rate, Chance Thayer, John E. Gilley, Lisa M. Durso, David B. Marx

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Manure is applied to cropland areas with varying surface cover to meet single‐year or multiple‐year crop nutrient requirements. The objectives of this field study were to: (1) examine runoff water quality characteristics following land application of manure to sites with and without wheat residue, (2) compare the water quality impacts of land application of manure to meet 0‐, 1‐, 2‐, 4‐, and 8‐year P‐based requirements for corn, and (3) evaluate the effects of varying runoff rates on runoff nutrient loads. Three 30‐min simulated rainfall events, separated by 24 h intervals, were applied at an intensity of 70 mm h^{‐1 …}

Field Application Uniformity And Accuracy Of Two Rate Control Systems With Automatic Section Capabilities On Agricultural Sprayers, Ajay Sharda, Joe D. Luck, John P. Fulton, Timothy P. Mcdonald, Scott A. Shearer

Department of Agricultural and Biological Systems Engineering: Faculty Publications

The adoption of automatic section control (ASC) on agricultural sprayers remains popular since it reduces overlap and application in unwanted areas leading to input savings and improved environmental stewardship. Most spray controllers attempt to maintain the desired target rate during ASC actuation (ON and OFF of control sections which change the width of boom-section actually spraying) but limited knowledge exists regarding controller response and nozzle discharge variation during field operation. Therefore, field experiments were conducted using two common self-propelled sprayers equipped with commercially available control systems with ASC capabilities. Pressure transducers were mounted across the spray booms to record real-time …

Value-Added Uses For Crude Glycerol–A Byproduct Of Biodiesel Production, Fangxia Yang, Milford Hanna, Runcang Sun

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Biodiesel is a promising alternative, and renewable, fuel. As its production increases, so does production of the principle co-product, crude glycerol. The effective utilization of crude glycerol will contribute to the viability of biodiesel. In this review, composition and quality factors of crude glycerol are discussed. The value-added utilization opportunities of crude glycerol are reviewed. The majority of crude glycerol is used as feedstock for production of other value-added chemicals, followed by animal feeds.

Quantification And Heterogeneity Of Infiltration And Transport In Alluvial Floodplains, Derek M. Heeren, Garey A. Fox, Daniel E. Storm, Peter Q. Storm, Brian E. Haggard, Todd Halihan, Ronald B. Miller

Department of Agricultural and Biological Systems Engineering: Faculty Publications

In order to protect drinking water systems and aquatic ecosystems, all critical nutrient source areas and transport mechanisms need to be characterized. It is hypothesized that hydrologic heterogeneities (e.g., macropores and gravel outcrops) in the subsurface of floodplains play an integral role in impacting flow and contaminant transport between the soil surface and shallow alluvial aquifers which are intricately connected to streams. Infiltration is often assumed to be uniform at the field scale, but this neglects the high spatial variability common in anisotropic, heterogeneous alluvial floodplain soils. In the Ozark ecoregion, for example, the erosion of carbonate bedrock (primarily limestone) …

Berm Method For Quantification Of Infiltration And Leaching At The Plot Scale In High Conductivity Soils, Derek M. Heeren, Garey A. Fox, Daniel E. Storm

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Measuring infiltration and leaching at the plot scale is difficult, especially for high hydraulic conductivity soils. Infiltration rate has been indirectly calculated at the plot scale by comparing surface runoff to rainfall. Direct measurement of infiltration and leaching beyond the point scale is typically limited to locations where land forming has been performed, e.g. infiltration ponds and fields with basin irrigation. The standard method for field measurement of infiltration is a double ring infiltrometer, which is limited in size (typically 30 cm diameter). In this research, a new method is proposed that uses a temporary berm constructed of a water …

Comparison Of 2-Way Versus Metered 3-Way Boom Shut-Off Valves For Automatic Section Control On Agricultural Sprayers, Ajay Sharda, Joe D. Luck, John P. Fulton, Timothy P. Mcdonald, Scott A. Shearer, Daniel K. Mullenix

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Modern spray rate controllers along with technologies such as automatic section control (ASC) provide benefits such as overlap reduction on agricultural sprayers. However, product (liquid) dynamics within the boom plumbing affect off-rate errors and application uniformity during rate changes and ASC actuation. Therefore, this study was conducted to compare nozzle flow stability and uniformity across the boom when using two different boom shut-off valves (2-way and metered 3-way) on an 18.3-m sprayer boom. Pressure transducers were mounted at 1) the boom manifold, 2) randomly at 12 nozzle bodies across the spray boom, and 3) upstream and downstream of the flow …

Selecting Sprinkler Packages For Center Pivots, Derrel L. Martin, William L. Kranz, Allen L. Thompson, Hong Liang

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Center pivots are the primary method of irrigation across the U.S. Great Plains. Center-pivot irrigation is also the fastest growing method of irrigation in the U.S. and around the world. Pivots have the potential to be very efficient and uniform if sprinkler devices are properly selected for local field conditions. New water application devices provide for selection that minimizes runoff and controls droplet sizes to reduce evaporation and drift losses. We present updates to models for computing runoff potential based on characteristics of sprinkler devices and soil textural classes. A dimensionless solution to the Green-Ampt infiltration method for center pivots …

Sensitivity Of Grass- And Alfalfa-Reference Evapotranspiration To Weather Station Sensor Accuracy, Dana O. Porter, Prasanna H. Gowda, Thomas H. Marek, Terry A. Howell, Jerry Moorhead, Suat Irmak

Department of Agricultural and Biological Systems Engineering: Faculty Publications

A sensitivity analysis was conducted to determine the relative effects of measurement errors in climate data input parameters on the accuracy of calculated reference crop evapotranspiration (ET) using the ASCE-EWRI Standardized Reference ET Equation. Data for the period of 1995 to 2008 from an automated weather station located at the USDA-ARS Conservation and Production Research Laboratory at Bushland, Texas were used for the analysis. Results

indicated that grass (ET_os) and alfalfa (ET_rs) reference crop ET were most sensitive to measurement errors in wind speed and air temperature followed by incoming shortwave (solar) radiation, and that data …

Evaluation Of A Rhodamine-Wt Dye/Glycerin Mixture As A Tracer For Testing Direct Injection Systems For Agricultural Sprayers, Joe D. Luck, Scott A. Shearer, Brian D. Luck, Fred A. Payne

Department of Agricultural and Biological Systems Engineering: Faculty Publications

The purpose of this study was to provide valuable insight regarding the use of Rhodamine WT (red) dye as a tracer for evaluating injected concentrations. More specifically, the effects of mixing the dye with glycerin to simulate the viscosity of a pesticide (e.g., glyphosate) or injecting the dye/glycerin mixture into deionized (DI) versus tap water on developing appropriate calibration equations were evaluated. Test results indicated that mixing the dye in a solution of glycerin and DI water significantly affected absorbance measurements compared to the dye mixed solely in DI water. The error in estimating absorbance was 7.4% between the two …

Swat: Model Use, Calibration, And Validation, Jeffrey G. Arnold, Daniel N. Moriasi, Philip W. Gassman, Karim C. Abbaspour, Michael J. White, Raghavan Srinivasan, Chinnasamy Santhi, Daren Harmel, Ann Van Griensven, Michael W. Van Liew, Narayanan Kannan, Manoj K. Jha

Department of Agricultural and Biological Systems Engineering: Faculty Publications

SWAT (Soil and Water Assessment Tool) is a comprehensive, semi-distributed river basin model that requires a large number of input parameters, which complicates model parameterization and calibration. Several calibration techniques have been developed for SWAT, including manual calibration procedures and automated procedures using the shuffled complex evolution method and other common methods. In addition, SWAT-CUP was recently developed and provides a decision-making framework that incorporates a semi-automated approach (SUFI2) using both manual and automated calibration and incorporating sensitivity and uncertainty analysis. In SWAT-CUP, users can manually adjust parameters and ranges iteratively between autocalibration runs. Parameter sensitivity analysis helps focus the …

Soil Water Extraction Patterns And Crop, Irrigation, And Evapotranspiration Water Use Efficiency Of Maize Under Full And Limited Irrigation And Rainfed Settings, Koffi Djaman, Suat Irmak

Department of Agricultural and Biological Systems Engineering: Faculty Publications

The effects of full and limited irrigation and rainfed maize production practices on soil water extraction and water use efficiencies were investigated in 2009 and 2010 under center-pivot irrigation near Clay Center, Nebraska. Four irrigation regimes (fully irrigated treatment (FIT), 75% FIT, 60% FIT, and 50% FIT) and a rainfed treatment were implemented. The crop water use efficiency (CWUE, or crop water productivity), irrigation water use efficiency (IWUE), and evapotranspiration water use efficiency (ETWUE) were used to evaluate the water productivity performance of each

treatment. The seasonal rainfall amounts in 2009 and 2010, respectively, were 426 mm (18% below normal) …

Large-Scale On-Farm Implementation Of Soil Moisture-Based Irrigation Management Strategies For Increasing Maize Water Productivity, Suat Irmak, Michael J. Burgert, Haishun Yang, Kenneth G. Cassman, Daniel T. Walters, William R. Rathje, Jose O. Payero, Patricio Grassini, Mark S. Kuzila, Kelly J. Brunkhorst, Dean E. Eisenhauer, William L. Kranz, Brandy Vandewalle, Jennifer M. Rees, Gary L. Zoubek, Charles A. Shapiro, Gregory J. Teichmeier

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Irrigated maize is produced on about 3.5 Mha in the U.S. Great Plains and western Corn Belt. Most irrigation water comes from groundwater. Persistent drought and increased competition for water resources threaten long-term viability of groundwater resources, which motivated our research to develop strategies to increase water productivity without noticeable reduction in maize yield. Results from previous research at the University of Nebraska-Lincoln (UNL) experiment stations in 2005 and 2006 found that it was possible to substantially reduce irrigation amounts and increase irrigation water use efficiency (IWUE) and crop water use efficiency (CWUE) (or crop water productivity) with little or …

Evaporative Losses From A Common Reed-Dominated Peachleaf Willow And Cottonwood Riparian Plant Community, Isa Kabenge, Suat Irmak

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Our study is one of the first to integrate and apply within-canopy radiation physics parameters and scaling-up leaf-level stomatal resistace (r_L) to canopy resistance (r_c) approach to quantify hourly transpiration (TRP) rates of individual riparian plant species—common reed (Phragmites australis), peachleaf willow (Salix amygdaloides), and cottonwood (Populus deltoides)— in a mixed riparian plant community in the Platte River Basin in central Nebraska. Two experimental years (2009 and 2010) were contrasted by warmer air temperature and presence of flood water in 2010. The seasonal average r_c values for common reed, …

Curcumin-Loaded Magnetic Nanoparticles For Breast Cancer Therapeutics And Imaging Applications, Murali M. Yallapu, Shadi F. Othman, Evan T. Curtis, Nichole A. Bauer, Neeraj Chauhan, Deepak Kumar, Meena Jaggi, Subhash C. Chauhan

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Breast cancer is a commonly diagnosed cancer among women in the United States. It is estimated that 230,480 women were diagnosed with, and 39,520 women died from, breast cancer in 2011 in the United States.¹ Basal-like breast cancer accounts for 15% of all breast cancers and has a poor prognosis. The majority of these cancers are referred to as triple-negative breast cancers because they do not over express estrogen receptor, progesterone receptor, or human epidermal growth factor receptor 2.^2,3 Chemotherapy is an effective option to treat such cancers; however, chemotherapy is always associated with several sets of side …

Do Clark’S Nutcrackers Demonstrate What-Where-When Memory On A Cache-Recovery Task?, Kristy L. Gould, Amy J. Ort, Alan C. Kamil

Avian Cognition Papers

What-where-when (WWW) memory during cache recovery was investigated in six Clark’s nutcrackers. During caching, both red- and blue-colored pine seeds were cached by the birds in holes filled with sand. Either a short (3 day) retention interval (RI) or a long (9 day) RI was followed by a recovery session during which caches were replaced with either a single seed or wooden bead depending upon the color of the cache and length of the retention interval. Knowledge of what was in the cache (seed or bead), where it was located, and when the cache had been made (3 or 9 …

High-Efficiency Talen-Based Gene Editing Produces Disease-Resistant Rice, Ting Li, Martin H. Spalding, Donald P. Weeks, Bing Yang

Department of Biochemistry: Faculty Publications

Transcription activator–like (TAL) effectors of Xanthomonas oryzae pv. oryzae (Xoo) contribute to pathogen virulence by transcriptionally activating specific rice disease-susceptibility (S) genes. TAL effector nucleases (TALENs)—fusion proteins derived from the DNA recognition repeats of native or customized TAL effectors and the DNA cleavage domains of FokI—have been used to create site-specific gene modifications in plant cells, yeast, animals, and even human pluripotent cells. Here, we exploit TALEN technology to edit a specific S gene in rice to thwart the virulence strategy of X. oryzae and thereby engineer heritable genome modifications for resistance to bacterial blight, a devastating disease …

Dengue Virus Infection Perturbs Lipid Homeostasis In Infected Mosquito Cells, Rushika Perera, Catherine Riley, Giorgis Isaac, Amber S. Hopf-Jannasch, Ronald J. Moore, Karl W. Weitz, Ljiljana Pasa-Tolic, Thomas O. Metz, Jiri Adamec, Richard J. Kuhn

Department of Biochemistry: Faculty Publications

Dengue virus causes ~50–100 million infections per year and thus is considered one of the most aggressive arthropodborne human pathogen worldwide. During its replication, dengue virus induces dramatic alterations in the intracellular membranes of infected cells. This phenomenon is observed both in human and vector-derived cells. Using high-resolution mass spectrometry of mosquito cells, we show that this membrane remodeling is directly linked to a unique lipid repertoire induced by dengue virus infection. Specifically, 15% of the metabolites detected were significantly different between DENV infected and uninfected cells while 85% of the metabolites detected were significantly different in isolated replication complex …

Udp-Glucose Dehydrogenase Polymorphisms From Patients With Congenital Heart Valve Defects Disrupt Enzyme Stability And Quaternary Assembly, Annastasia S. Hyde, Erin L. Farmer, Katherine E. Easley, Kristy Van Lammeren, Vincent M. Christoffels, Joseph J. Barycki, Jeroen Bakkers, Melanie A. Simpson

Department of Biochemistry: Faculty Publications

Background: UDP-glucose dehydrogenase (UGDH) polymorphisms were identified in a screen of candidate genes for heart valve defects.

Results: Two individual mutants fail to rescue cardiac valve defects in UGDH-deleted zebrafish and have reduced stability in vitro.

Conclusion: UGDH loss of function mutations result in a subset of human congenital cardiac valve defects caused by reduced enzyme activity during morphogenesis.

Significance: Screening these alleles could predict valve defects.

Genome, Functional Gene Annotation, And Nuclear Transformation Of The Heterokont Oleaginous Alga Nannochloropsis Oceanica Ccmp1779, Astrid Vieler, Guangxi Wu, Chia-Hong Tsai, Blair Bullard, Adam J. Cornish, Christopher Harvey, Ida-Barbara Reca, Chelsea Thornburg, Rujira Achawanantakun, Christopher J. Buehl, Michael S. Campbell, David Cavalier, Kevin L. Childs, Teresa J. Clark, Rahul Deshpande, Erika Erickson, Ann Armenia Ferguson, Witawas Handee, Que Kong, Xiaobo Li, Bensheng Liu, Steven Lundback, Cheng Peng, Rebecca Roston, Michigan State University, Jeffrey P. Simpson, Allan Terbush, Jaruswan Warakanont, Simone Zäuner, Eva M. Farre, Eric L. Hegg, Ning Jiang, Min-Hao Kuo, Yan Lu, Krishna K. Niyogi, John Ohlrogge, Katherine W. Osteryoung, Yair Shachar-Hill, Barbara B. Sears, Yanni Sun, Hideki Takahashi, Mark Yandell, Shin-Han Shiu, Christoph Benning

Department of Biochemistry: Faculty Publications

Unicellular marine algae have promise for providing sustainable and scalable biofuel feedstocks, although no single species has emerged as a preferred organism. Moreover, adequate molecular and genetic resources prerequisite for the rational engineering of marine algal feedstocks are lacking for most candidate species. Heterokonts of the genus Nannochloropsis naturally have high cellular oil content and are already in use for industrial production of high-value lipid products. First success in applying reverse genetics by targeted gene replacement makes Nannochloropsis oceanica an attractive model to investigate the cell and molecular biology and biochemistry of this fascinating organism group. Here we present the …

Cellular Responses Of Candida Albicans To Phagocytosis And The Extracellular Activities Of Neutrophils Are Critical To Counteract Carbohydrate Starvation, Oxidative And Nitrosative Stress, Pedro Miramón, Christine Dunker, Hanna Windecker, Iryna Bohovych, Alistair J.P. Brown, Oliver Kurzai, Bernhard Hube

Department of Biochemistry: Faculty Publications

Neutrophils are key players during Candida albicans infection. However, the relative contributions of neutrophil activities to fungal clearance and the relative importance of the fungal responses that counteract these activities remain unclear. We studied the contributions of the intra- and extracellular antifungal activities of human neutrophils using diagnostic Green Fluorescent Protein (GFP)-marked C. albicans strains. We found that a carbohydrate starvation response, as indicated by upregulation of glyoxylate cycle genes, was only induced upon phagocytosis of the fungus. Similarly, the nitrosative stress response was only observed in internalised fungal cells. In contrast, the response to oxidative stress was observed in …

Small But Crucial: The Novel Small Heat Shock Protein Hsp21 Mediates Stress Adaptation And Virulence In Candida Albicans, Francois L. Mayer, Duncan Wilson, Ilse D. Jacobsen, Pedro Miramón, Silvia Slesiona, Iryna Bohovych, Alistair J.P. Brown, Bernhard Hube

Department of Biochemistry: Faculty Publications

Small heat shock proteins (sHsps) have multiple cellular functions. However, the biological function of sHsps in pathogenic microorganisms is largely unknown. In the present study we identified and characterized the novel sHsp Hsp21 of the human fungal pathogen Candida albicans. Using a reverse genetics approach we demonstrate the importance of Hsp21 for resistance of C. albicans to specific stresses, including thermal and oxidative stress. Furthermore, a hsp21∆/∆ mutant was defective in invasive growth and formed significantly shorter filaments compared to the wild type under various filamentinducing conditions. Although adhesion to and invasion into human-derived endothelial and oral epithelial cells …

Oligomerization Of Heme O Synthase In Cytochrome Oxidase Biogenesis Is Mediated By Cytochrome Oxidase Assembly Factor Coa2, Oleh Khalimonchuk, Hyung Kim, Talina Watts, Xochitl Perez-Martinez, Dennis R. Winge

Department of Biochemistry: Faculty Publications

The synthesis of the heme a cofactor used in cytochrome c

oxidase (CcO) is dependent on the sequential action of heme o

synthase (Cox10) and heme a synthase (Cox15). The active state of

Cox10 appears to be a homo-oligomeric complex,andformation of

this complex is dependent on the newly synthesized CcO subunit

Cox1 and the presence of an early Cox1 assembly intermediate.

Cox10 multimerization is triggered by progression of Cox1 from

the early assembly intermediate todownstreamintermediates.The

CcOassembly factor Coa2 appears important in coupling the presence

of newly synthesized Cox1 to Cox10 oligomerization. Cells

lacking Coa2 are impaired in Cox10 complex formation …

Selective Oma1 Protease-Mediated Proteolysis Of Cox1 Subunit Of Cytochrome Oxidase In Assembly Mutants, Oleh Khalimonchuk, Mi-Young Jeong, Talina Watts, Elliott Ferris, Dennis R. Winge

Department of Biochemistry: Faculty Publications

Background: Yeast lacking Coa2 are deficient in cytochrome c oxidase due to Cox1 degradation.

Results: Oma1 mediates Cox1 degradation in coa2∆ cells but not other mutants stalled in oxidase biogenesis.

Conclusion: Impaired hemylation of Cox1 in coa2∆ cells leads to misfolding and facile degradation by Oma1.

Significance: Oma1 functions in quality control of cytochrome oxidase assembly.

The Evolutionary Rewiring Of Ubiquitination Targets Has Reprogrammed The Regulation Of Carbon Assimilation In The Pathogenic Yeast Candida Albicans, Doblin Sandai, Zhikang Yin, Laura Selway, David Stead, Janet Walker, Michelle D. Leach, Iryna Bohovych, Iuliana V. Ene, Stavroula Kastora, Susan Budge, Carol A. Munro, Frank C. Odds, Neil A.R. Gow, Alistair J.P. Brown

Department of Biochemistry: Faculty Publications

Microbes must assimilate carbon to grow and colonize their niches. Transcript profiling has suggested that Candida albicans, a major pathogen of humans, regulates its carbon assimilation in an analogous fashion to the model yeast Saccharomyces cerevisiae, repressing metabolic pathways required for the use of alterative nonpreferred carbon sources when sugars are available. However, we show that there is significant dislocation between the proteome and transcriptome in C. albicans. Glucose triggers the degradation of the ICL1 and PCK1 transcripts in C. albicans, yet isocitrate lyase (Icl1) and phosphoenolpyruvate carboxykinase (Pck1) are stable and are retained. Indeed, numerous …

Role Of Apoptosis-Inducing Factor, Proline Dehydrogenase, And Nadph Oxidase In Apoptosis And Oxidative Stress, Sathish Kumar Natarajan, Donald F. Becker

Department of Biochemistry: Faculty Publications

Flavoproteins catalyze a variety of reactions utilizing flavin mononucleotide or flavin adenine dinucleotide as cofactors. The oxidoreductase properties of flavoenzymes implicate them in redox homeostasis, oxidative stress, and various cellular processes, including programmed cell death. Here we explore three critical flavoproteins involved in apoptosis and redox signaling, ie, apoptosis-inducing factor (AIF), proline dehydrogenase, and NADPH oxidase. These proteins have diverse biochemical functions and influence apoptotic signaling by unique mechanisms. The role of AIF in apoptotic signaling is two-fold, with AIF changing intracellular location from the inner mitochondrial membrane space to the nucleus upon exposure of cells to apoptotic stimuli. In …

Late-Stage Maturation Of The Rieske Fe/S Protein: Mzm1 Stabilizes Rip1 But Does Not Facilitate Its Translocation By The Aaa Atpase Bcs1, Tie-Zhong Cui, Pamela M. Smith, Jennifer L. Fox, Oleh Khalimonchuk, Dennis R. Winge

Department of Biochemistry: Faculty Publications

The final step in the assembly of the ubiquinol-cytochrome c reductase or bc₁ complex involves the insertion of the Rieske Fe/S cluster protein, Rip1. Maturation of Rip1 occurs within the mitochondrial matrix prior to its translocation across the inner membrane (IM) in a process mediated by the Bcs1 ATPase and subsequent insertion into the bc₁ complex. Here we show that the matrix protein Mzm1 functions as a Rip1 chaperone, stabilizing Rip1 prior to the translocation step. In the absence of Mzm1, Rip1 is prone to either proteolytic degradation or temperature-induced aggregation. A series of Rip1 truncations were engineered …

Substrate Channeling In Proline Metabolism, Benjamin W. Arentson, Nikhilesh Sanyal, Donald F. Becker

Department of Biochemistry: Faculty Publications

Proline metabolism is an important pathway that has relevance in several cellular functions such as redox balance, apoptosis, and cell survival. Results from different groups have indicated that substrate channeling of proline metabolic intermediates may be a critical mechanism. One intermediate is pyrroline-5-carboxylate (P5C), which upon hydrolysis opens to glutamic semialdehyde (GSA). Recent structural and kinetic evidence indicate substrate channeling of P5C/ GSA occurs in the proline catabolic pathway between the proline dehydrogenase and P5C dehydrogenase active sites of bifunctional proline utilization A (PutA). Substrate channeling in PutA is proposed to facilitate the hydrolysis of P5C to GSA which is …

Glutaredoxin 1 Protects Dopaminergic Cells By Increased Protein Glutathionylation In Experimental Parkinson's Disease, Humberto Rodriguez-Rocha, Aracely Garcia Garcia, Laura Zavala-Flores, Sumin Li, Nandakumar Madayiputhiya, Rodrigo Franco

Department of Biochemistry: Faculty Publications

Aims: Chronic exposure to environmental toxicants, such as paraquat, has been suggested as a risk factor for Parkinson's disease (PD). Although dopaminergic cell death in PD is associated with oxidative damage, the molecular mechanisms involved remain elusive. Glutaredoxins (GRXs) utilize the reducing power of glutathione to modulate redox-dependent signaling pathways by protein glutathionylation. We aimed to determine the role of GRX1 and protein glutathionylation in dopaminergic cell death. Results: In dopaminergic cells, toxicity induced by paraquat or 6-hydroxydopamine (6-OHDA) was inhibited by GRX1 overexpression, while its knock-down sensitized cells to paraquat-induced cell death. Dopaminergic cell death was paralleled by protein …