Mechanisms Of Translational Repression Of The Sperm Mitochondria Associated-Cysteine Rich Protein (Smcp) Mrna In Round Spermatids, Danielle L. Cullinane

Graduate Doctoral Dissertations

The sperm-mitochondria-associated cysteine-rich protein (SMCP) is a male germ cell-specific protein that localizes to the outer membranes of sperm mitochondria and increases sperm motility. The Smcp mRNA is transcribed in early spermatids, and stored in a translationally repressed state for ~7 days before translation is activated in late spermatids. Identifying the cis-elements and trans-factors that repress the Smcp mRNA in early spermatids is important because these factors and elements coordinate the translational activity of hundreds of mRNAs.

A mutation was studied in transgenic mice in which the 16 nucleotides downstream of the first poly(A) signal in the Smcp …

Amylin Structure, Aggregation, And Pancreatic Β Cell Toxicity, Sharadrao Patil

MCB Articles

In most type 2 diabetes patients, amyloid plaques have been found juxtaposed with membranes of pancreatic β-cells. These plaques are composed of amyloid fibrils of the 37 residue endocrine hormone amylin and cause distinct changes in cell membrane morphology associated with the destruction of β-cells. Research is still ongoing to identify the toxic species involved and the mechanisms by which mature fibrils or oligomers cause cytotoxicity. The projects undertaken were designed to study the molecular structural features of amylin, mechanism of amyloid aggregation and, to develop cytotoxicity inhibitors. We determined the structure of human amylin bound to SDS micelles using …

Real-Time Kinetic Studies Of Bacillus Subtilis Oxalate Decarboxylase And Ceriporiopsis Subvermispora Oxalate Oxidase Using Luminescent Oxygen Sensor, Laura Molina, Thomas Goodall, Umar Twahir, Ellen W. Moomaw

Faculty and Research Publications

Oxalate decarboxylase (OxDC), an enzyme of the bicupin superfamily, catalyzes the decomposition of oxalate into carbon dioxide and formate at an optimal pH of 4.3 in the presence of oxygen. However, about 0.2% of all reactions occur through an oxidase mechanism that consumes oxygen while producing two equivalents of carbon dioxide and one equivalent of hydrogen peroxide. The kinetics of oxidase activity were studied by measuring the consumption of dissolved oxygen over time using a luminescent oxygen sensor. We describe the implementation of and improvements to the oxygen consumption assay. The oxidase activity of wild type OxDC was compared to …

Epigenetic Regulation Of Nuclear Hormone Receptor Dax-1, Michael B. Heskett

Master's Theses

DAX-1 (NR0B1) is an orphan nuclear receptor that plays a key role in the development and maintenance of steroidogenic tissue in mammals. Dax-1 is also expressed in mouse embryonic stem (ES) cells and is required to maintain pluripotency. Duplication of the X-chromosome in the region containing the NR0B1 gene results in sex reversal, and mutations in NR0B1 cause adrenal hypoplasia congenita. DAX-1 has been observed to act as a corepressor of other nuclear receptors including androgen receptor (AR), estrogen receptor (ER), and steroidogenic factor 1 (SF-1). In addition to pluripotent ES cells, DAX-1 is primarily expressed in select tissues of …

Mtorc2 Is Required For Rit-Mediated Oxidative Stress Resistance, Weikang Cai, Douglas A. Andres

Molecular and Cellular Biochemistry Faculty Publications

Rit, a member of the Ras family of GTPases, has been shown to promote cell survival in response to oxidative stress, in part by directing an evolutionarily conserved p38 MAPK-Akt survival cascade. Aberrant Rit signaling has recently been implicated as a driver mutation in human cancer, adding importance to the characterization of critical Rit effector pathways. However, the mechanism by which Rit-p38 signaling regulated Akt activity was unknown. Here, we identify mTORC2 as a critical downstream mediator of Rit-dependent survival signaling in response to reactive oxygen species (ROS) stress. Rit interacts with Sin1 (MAPKAP1), and Rit loss compromises ROS-dependent mTORC2 …

Investigating The Links Between Ozone And Organic Aerosol Chemistry In A Biomass Burning Plume From A Prescribed Fire In California Chaparral, M. J. Alvarado, C. R. Lonsdale, Robert Yokelson, S. K. Akagi, H. Coe, J. S. Craven, E. V. Fischer, G. R. Mcmeeking, J. H. Seinfeld, T. Soni, J. W. Taylor, D. R. Weise, C. E. Wold

Chemistry and Biochemistry Faculty Publications

Within minutes after emission, rapid, complex photochemistry within a biomass burning smoke plume can cause large changes in the concentrations of ozone (O₃) and organic aerosol (OA). Being able to understand and simulate this rapid chemical evolution under a wide variety of conditions is a critical part of forecasting the impact of these fires on air quality, atmospheric composition, and climate. Here we use version 2.1 of the Aerosol Simulation Program (ASP) to simulate the evolution of O₃ and secondary organic aerosol (SOA) within a young biomass burning smoke plume from the Williams prescribed burn in chaparral, …

Fap206 Is A Microtubule-Docking Adapter For Ciliary Radial Spoke 2 And Dynein C, Krishna K. Vasudevan, Kangkang Song, Lea M. Alford, Winfield Sale, Erin E. Dymek, Elizabeth F. Smith, Todd Hennessey, Ewa Joachimiak, Paulina Urbanska, Dorota Wloga, William Dentler, Daniela Nicastro, Jacek Gaertig

Dartmouth Scholarship

Radial spokes are conserved macromolecular complexes that are essential for ciliary motility. A triplet of three radial spokes, RS1, RS2, and RS3, repeats every 96 nm along the doublet microtubules. Each spoke has a distinct base that docks to the doublet and is linked to different inner dynein arms. Little is known about the assembly and functions of individual radial spokes. A knockout of the conserved ciliary protein FAP206 in the ciliate Tetrahymena resulted in slow cell motility. Cryo–electron tomography showed that in the absence of FAP206, the 96-nm repeats lacked RS2 and dynein c. Occasionally, RS2 assembled but lacked …

A Microrna-1280/Jag2 Network Comprises A Novel Biological Target In High-Risk Medulloblastoma, Fengfei Wang, Marc Remke, Tze-Chen Hsieh, Lizi Wu, Cynthia Hawkins, Joseph M. Wu, Erxi Wu

NYMC Faculty Publications

Over-expression of PDGF receptors (PDGFRs) has been previously implicated in high-risk medulloblastoma (MB) pathogenesis. However, the exact biological functions of PDGFRα and PDGFRβ signaling in MB biology remain poorly understood. Here, we report the subgroup specific expression of PDGFRα and PDGFRβ and their associated biological pathways in MB tumors. c-MYC, a downstream target of PDGFRβ but not PDGFRα, is involved in PDGFRβ signaling associated with cell proliferation, cell death, and invasion. Concurrent inhibition of PDGFRβ and c-MYC blocks MB cell proliferation and migration synergistically. Integrated analysis of miRNA and miRNA targets regulated by both PDGFRβ and c-MYC reveals that increased …

Regulation Of Phialide Morphogenesis In Aspergillus Nidulans, Hu Yin

School of Biological Sciences: Dissertations, Theses, and Student Research

Filamentous fungi have two distinctive life cycles, vegetative growth and development for sexual or asexual spore formation. The asexual reproduction in development as conidiation in A. nidulans is the dominant form of producing spores effectively. A complex conidiophore structure is developed during asexual reproduction process. The conidiophore is formed from hyphal cell and consists of stalk, vesicle, metulae, phialide and conidial spores. Phialides are essential sporogenous cells in the conidiophore structure. The growth pattern is switched from acropetal to basipetal between phialide and spores, which makes phialide a unique cell type in A. nidulans and other phialide producing fungi. Study …

Biological And Physiological Condition Of Juvenile California Halibut (Paralichthys Californicus) Exposed To A Contamination Gradient In Mission Bay, Ca, Kevin Stolzenbach

Theses

Contaminated sediments in marine environments have been shown to be good indicators of ecological risk and a means to assess anthropogenic impacts on marine habitats and the animals that inhabit them (Long et al. 1995, Rattner 2009). Estuarine sediments are especially complex media with regard to physical, chemical, and biological characteristics that trap, store, modify and sometimes release contaminants to the biota (Long et al. 1995). Especially vulnerable are animals that are in constant contact with the sediments, such as flatfishes that partially bury themselves for camouflage (Costa et al. 2011). Impacts can be assessed in a number of ways, …

Development Of An Immunodiagnostic Assay For Canine Tear Lacritin, Alison Mae Enghauser

Senior Honors Projects, 2010-2019

Purpose: Lacritin is a naturally occurring human glycoprotein secreted from the lacrimal gland as a component of tears. Preliminary studies suggest that down-regulation of lacritin is associated with various ocular diseases, such as dry eye syndrome and blepharitis. Thus, lacritin shows potential as a new topical therapeutic for the treatment of ocular diseases. Previous studies have shown that recombinant human lacritin, when topically applied to rabbit eyes, promotes basal tearing. Antibodies produced against the terminal ends of human lacritin were used to develop a clinical immunodiagnostic assay that detect and quantify lacritin in human tear samples. In order to develop …

Synthesis And Characterization Of Polymer (Sulfonated Poly-Ether-Ether-Ketone) Based Nanocomposite (H-Boron Nitride) Membrane For Hydrogen Storage, Naresh Muthu, S. Rajashabala, R. Kannan

Chemistry and Biochemistry Faculty Publications

The development of light weight and compact hydrogen storage materials is still prerequisite to fuel-cell technology to be fully competitive. The present experimental study reports the hydrogen storage capability of sulfonated poly-ether-ether-ketone (SPEEK)-hexagonal boron nitride (h-BN) (SPEEK-h-BN) nanocomposite membranes. The nanocomposite membranes are prepared by considering various amount of h-BN (0, 1, 3 and 5 wt. %) by phase inversion technique. The degree of sulfonation of the PEEK (SPEEK) is found to be 65% by Proton Nuclear Magnetic Resonance (¹H NMR) spectroscopy. Hydrogen adsorption studies have been carried out using a Seiverts-like hydrogenation setup. The membranes are characterized …

Loss Of Thiol Repair Systems In Human Cataractous Lenses, Min Wei, Kui-Yi Xing, Yin-Chuan Fan, Teodosio Libondi, Marjorie F. Lou

School of Veterinary and Biomedical Sciences: Faculty Publications

PURPOSE. The purpose of this study was to investigate the thiol repair systems of thioltransferase (TTase) and thioredoxin (Trx) and oxidation-damaged proteins in human cataractous lenses.

METHODS. Cataractous lenses in humans (57–85 years of age) were classified into cortical, nuclear, mixed, mature, and hypermature cataract types by using a lens opacity classification system, and were obtained by extracapsular cataract extraction (ECCE) procedure. Cortical and nuclear cataracts were grouped by decreasing order of visual acuity into optical chart reading (R), counting fingers (CF), hand motion (HM), and light perception (LP). ECCE lens homogenate was analyzed for glutathione (GSH) level and enzyme …

Transcriptomic And Proteomic Dynamics In The Metabolism Of A Diazotrophic Cyanobacterium, Cyanothece Sp. Pcc 7822 During A Diurnal Light-Dark Cycle, Louis Sherman

Louis A Sherman

Background: Cyanothece sp. PCC 7822 is an excellent cyanobacterial model organism with great potential to be applied as a biocatalyst for the production of high value compounds. Like other unicellular diazotrophic cyanobacterial species, it has a tightly regulated metabolism synchronized to the light-dark cycle. Utilizing transcriptomic and proteomic methods, we were able to quantify the relationships between transcription and translation underlying central and secondary metabolism in response to nitrogen free, 12 hour light and 12 hour dark conditions.

Results: By combining mass-spectrometry based proteomics and RNA-sequencing transcriptomics, we quantitatively measured a total of 6766 mRNAs and 1322 proteins at four …

Effect Of Hydrogen Peroxide On The Biosynthesis Of Heme And Proteins: Potential Implications For The Partitioning Of Glu-TrnaGlu Between These Pathways, Carolina Farah, Gloria Levicán, Michael Ibba, Omar Orellana

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Glutamyl-tRNA (Glu-tRNA^Glu) is the common substrate for both protein translation and heme biosynthesis via the C5 pathway. Under normal conditions, an adequate supply of this aminoacyl-tRNA is available to both pathways. However, under certain circumstances, Glu-tRNA^Glu can become scarce, resulting in competition between the two pathways for this aminoacyl-tRNA. In Acidithiobacillus ferrooxidans, glutamyl-tRNA synthetase 1 (GluRS1) is the main enzyme that synthesizes Glu-tRNA^Glu. Previous studies have shown that GluRS1 is inactivated in vitro by hydrogen peroxide (H₂O₂). This raises the question as to whether H₂O₂ negatively affects …

Hcpe, A Potential Immuno-Modulatory Protein From Helicobacter Pylori That Is Dependent On The Disulfide Bond Protein Dsbhp, Jeff Lester

Electronic Thesis and Dissertation Repository

H. pylori is a human gastric pathogen that colonizes ~ 50% of the world’s population. It can cause gastritis, gastric or duodenal ulcers and also gastric cancer. H. pylori produces Helicobacter cysteine rich protein HcpE, a secreted protein which may play a role in virulence. In this study we show that HcpE is secreted in the culture supernatant both as a soluble protein and in association with outer membrane vesicles, and may play a role in the modulation of H. pylori inflammatory responses. We identified that Dsb_HP is necessary for HcpE production and secretion in H. pylori, and …

Dancing Through Life: Allosteric Transitions And Structural Analysis Of Hsp70 And Hsp110 Chaperone Proteins, Gabrielle Stetz, Gennady M. Verkhivker

Student Scholar Symposium Abstracts and Posters

The molecular chaperone protein Hsp70 is centrally involved in cellular homeostasis by assisting in the folding and degradation of protein substrates. Hsp70 is joined by co-chaperones, such as Hsp110, which contribute to specialized tasks of the Hsp70 complex. Imbalances of this heat shock protein system are believed to be involved with the deregulation of cancer pathways and other human diseases. Better understanding of how these heat shock proteins work at the molecular level, which has been investigated using molecular docking tools, will give more clues about biological function. Simulating the formation and function of Hsp70 based chaperone complexes could provide …

Argument Map: Loewi's Argument That Neuro-Transmission Works With Chemical Signals Instead Of Eletrical (Short Version), Michael Hoffmann

Michael H.G. Hoffmann

This argument shows how the hypothesis that muscles are probably stimulated exclusively by chemical signals and not by electrical ones can be justified by Loewi's experimentum crucis.

Effect Of Intracellular Expression Of Antimicrobial Peptide Ll-37 On Growth Of Escherichia Coli Strain Top10 Under Aerobic And Anaerobic Conditions, Wei Liu, Shi Dong, Fei Xu, Xue Wang, T. Withers, Hongwei Yu, Xin Wang

Hongwei Yu

Antimicrobial peptides (AMPs) can cause lysis of target bacteria by directly inserting themselves into the lipid bilayer. This killing mechanism confounds the identification of the intracellular targets of AMPs. To circumvent this, we used a shuttle vector containing the inducible expression of a human cathelicidin-related AMP, LL-37, to examine its effect on Escherichia coli TOP10 under aerobic and anaerobic growth conditions. Induction of LL-37 caused growth inhibition and alteration in cell morphology to a filamentous phenotype. Further examination of the E. coli cell division protein FtsZ revealed that LL-37 did not interact with FtsZ. Moreover, intracellular expression of LL-37 results …

Design, Assessment, And In Vivo Evaluation Of A Computational Model Illustrating The Role Of Cav1 In Cd4+ T-Lymphocytes, Brittany D. Conroy, Tyler A. Herek, Timonthy D. Shew, Matthew Latner, Joshua J. Larson, Laura Allen, Paul H. Davis, Tomáš Helikar, Christine E. Cutucache

Department of Biochemistry: Faculty Publications

Caveolin-1 (CAV1) is a vital scaffold protein heterogeneously expressed in both healthy and malignant tissue. We focus on the role of CAV1 when overexpressed in T-cell leukemia. Previously, we have shown that CAV1 is involved in cell-to-cell communication, cellular proliferation, and immune synapse formation; however, the molecular mechanisms have not been elucidated.We hypothesize that the role of CAV1 in immune synapse formation contributes to immune regulation during leukemic progression, thereby warranting studies of the role of CAV1 in CD4⁺ T-cells in relation to antigen-presenting cells. To address this need, we developed a computational model of a CD4⁺ immune …

Antifungal Amphiphilic Aminoglycoside K20: Bioactivities And Mechanism Of Action, Sanjib K. Shrestha, Cheng-Wei Tom Chang, Nicole Meissner, John Oblad, Jaya P. Shrestha, Kevin N. Sorensen, Michelle M. Grilley, Jon Y. Takemoto

Chemistry and Biochemistry Faculty Publications

K20 is a novel amphiphilic antifungal aminoglycoside that is synthetically derived from the antibiotic kanamycin A. Reported here are investigations of K20′s antimicrobial activities, cytotoxicity, and fungicidal mechanism of action. In vitro growth inhibitory activities against a variety of human and plant pathogenic yeasts, filamentous fungi, and bacteria were determined using microbroth dilution assays and time-kill curve analyses, and hemolytic and animal cell cytotoxic activities were determined. Effects on Cryptococcus neoformans H-99 infectivity were determined with a preventive murine lung infection model. The antifungal mechanism of action was studied using intact fungal cells, yeast lipid mutants, and small unilamellar lipid …

Odd-Even Effects In Charge Transport Across N-Alkanethiolate-Based Sams, Mostafa Baghbanzadeh, Felice C. Simeone, Carleen M. Bowers, Kung Ching Liao, Martin Thuo, Mahdi Baghbanzadeh, Michael S. Miller, Tricia Breen Carmichael, George M. Whitesides

Chemistry and Biochemistry Publications

This paper compares rates of charge transport across self-assembled monolayers (SAMs) of n-alkanethiolates having odd and even numbers of carbon atoms (nodd and neven) using junctions with the structure MTS/SAM//Ga2O3/EGaIn (M = Au or Ag). Measurements of current density, J(V), across SAMs of n-alkanethiolates on AuTS and AgTS demonstrated a statistically significant odd-even effect on AuTS, but not on AgTS, that could be detected using this technique. Statistical analysis showed the values of tunneling current density across SAMs of n-alkanethiolates on AuTS with nodd and neven belonging to two separate sets, and while there is a significant difference between the …

Gaip Interacting Protein C-Terminus Regulates Autophagy And Exosome Biogenesis Of Pancreatic Cancer Through Metabolic Pathways, Santanu Bhattacharya, Krishnendu Pal, Anil K. Sharma, Shamit K. Dutta, Julie S. Lau, Irene K. Yan, Enfeng Wang, Ahmed Elkhanany, Khalid M. Alkharfy, Arunik Sanyal, Tushar C. Patel, Suresh T. Chari, Mark R. Spaller, Debabrata Mukhopadhyay

Dartmouth Scholarship

GAIP interacting protein C terminus (GIPC) is known to play an important role in a variety of physiological and disease states. In the present study, we have identified a novel role for GIPC as a master regulator of autophagy and the exocytotic pathways in cancer. We show that depletion of GIPC-induced autophagy in pancreatic cancer cells, as evident from the upregulation of the autophagy marker LC3II. We further report that GIPC regulates cellular trafficking pathways by modulating the secretion, biogenesis, and molecular composition of exosomes. We also identified the involvement of GIPC on metabolic stress pathways regulating autophagy and microvesicular …

Understanding Ten-Eleven Translocation-2 In Hematological And Nervous Systems, Feng Pan

FIU Electronic Theses and Dissertations

I proposed the study of two distinct aspects of Ten-Eleven Translocation 2 (TET2) protein for understanding specific functions in different body systems.

In Part I, I characterized the molecular mechanisms of Tet2 in the hematological system. As the second member of Ten-Eleven Translocation protein family, TET2 is frequently mutated in leukemic patients. Previous studies have shown that the TET2 mutations frequently occur in 20% myelodysplastic syndrome/myeloproliferative neoplasm (MDS/MPN), 10% T-cell lymphoma leukemia and 2% B-cell lymphoma leukemia. Genetic mouse models also display distinct phenotypes of various types of hematological malignancies. I performed 5-hydroxymethylcytosine (5hmC) chromatin immunoprecipitation sequencing (ChIP-Seq) and RNA …

Novel Mechanisms In The Sorting Of Proglucagon To The Secretory Granules Of The Regulated Secretory Pathway, Leonard M. Guizzetti

Electronic Thesis and Dissertation Repository

The prohormone proglucagon encodes for multiple peptide hormones, including glucagon, glucagon-like peptide-1 (GLP-1), and GLP-2, produced through tissue-specific processing by prohormone convertase (PC) 1/3 and PC2. In alpha cells, PC2 yields glucagon, the major counter-regulatory hormone to insulin, which together, control glucose homeostasis. In contrast, GLP-1 and GLP2 are mainly produced in intestinal L-cells by PC1/3. GLP-1 stimulates insulin secretion following a meal, and therefore has opposing function to glucagon regulating glucose homeostasis; in contrast, GLP-2 enhances gut nutrient absorption. Efficient sorting of proglucagon to secretory granules is required for nutrient-regulated secretion. The aim of this thesis is to discover …

Ultrasmooth Gold Surfaces Prepared By Chemical Mechanical Polishing For Applications In Nanoscience, Michael S. Miller, Michael Anthony Ferrato, Adrian Niec, Mark C. Biesinger, Tricia Breen Carmichael

Chemistry and Biochemistry Publications

For over 20 years, template stripping has been the best method for preparing ultrasmooth metal surfaces for studies of nanostructures. However, the organic adhesives used in the template stripping method are incompatible with many solvents, limiting the conditions that may subsequently be used to prepare samples; in addition, the film areas that can be reliably prepared are typically limited to ∼1 cm2. In this article, we present chemical-mechanical polishing (CMP) as an adhesive-free, scalable method of preparing ultrasmooth gold surfaces. In this process, a gold film is first deposited by e-beam evaporation onto a 76-mm-diameter silicon wafer. The CMP process …

Melatonin And Neurogenesis: A Comparative Study Of The Efficacy Of Melatonin, Its Precursors, And L-Dopa On Neural Stem Cell Metabolism In Human Adult Neurospheres, Omar Heriba

HIM 1990-2015

Human neurosphere stem cells offer promising potential for the treatment of neurodegenerative diseases. Their well characterized multi-potency of differentiating into neurons, astrocytes, and oligodendrocytes when exposed to the optimum exogenous growth factors make them an exciting area of study (38). Finding novel endogenous methods of modulating stem cell metabolism will allow for the safer treatment of various brain disorders (34). In this experiment, melatonin, N-acetylserotonin, L-tryptophan, and L-DOPA are added in three different concentrations to neurospheres suspended in HNSC/GBM media with less than optimal concentrations of exogenous epidermal growth factor (EGF) and fibroblast growth factor (FGF). The alamarBlue assay (resazurin) …

Using The Yeast Two-Hybrid System To Determine The Function Of Parkin E3 Ubiquitin Ligase, Vanessa Nguyen

HIM 1990-2015

Parkin is a cytosolic E3 ubiquitin ligase that is recruited to the mitochondria during cellular stress and has been suggested to be involved in a variety of biological processes such as mitophagy. The recruitment of Parkin (PARK2) to the mitochondria is dependent upon the kinase activity and the accumulation of PINK1 on damaged mitochondria. Mutations in either PINK1 or Parkin genes disrupt this protective pathway and lead to the accumulation of damaged mitochondria. From a clinical standpoint, mutations in the PARK2 gene have been associated with the progression and onset of autosomal recessive juvenile parkinsonism. Without the presence of a …

The Dissolution Of Cellulose In Ionic Liquids - A Molecular Dynamics Study, Barmak Mostofian

Doctoral Dissertations

The use of ionic liquids for the dissolution of cellulose promises an alternative method for the thermochemical pretreatment of biomass that may be more efficient and environmentally acceptable than conventional techniques in aqueous solution. Understanding how ionic liquids act on cellulose is essential for improving pretreatment conditions and thus detailed knowledge of the interactions between solute and solvent molecules is necessary. Here, results from the first all-atom molecular dynamics simulation of an entire cellulose microfibril in 1-butyl-3-methylimidazolium chloride (BmimCl) are presented and the interactions and orientations of solvent ions with respect to glucose units on the hydrophobic and hydrophilic surfaces …

Pore Selectivity And Gating Of Arabidopsis Nodulin 26 Intrinsic Proteins And Roles In Boric Acid Transport In Reproductive Growth, Tian Li

Doctoral Dissertations

Plant nodulin-26 intrinsic proteins (NIPs) are members of the aquaporin superfamily that serve as multifunctional channels of uncharged metabolites and water. They share the same canonical hourglass fold as the aquaporin family. The aromatic arginine (ar/R) selectivity filter controls transport selectivity based on size, hydrophobicity, and hydrogen bonding with substrates. In Arabidopsis thaliana, NIP II subclass proteins contain a conserved ar/R “pore signature” that is composed of Alanine at the helix 2 position (H2), Valine/Isoleucine at the helix 5 position (H5), and an Alanine (LE1) and an invariant Arginine (LE2) at the two loop E positions. In this study, …