2009-2010 Projects
HFE: An Iron Uptake Regulation Molecule
Brookfield Central High School
Students: Yuhan Chen, Bryan Dongre, Justin Fu, Zach Gerner, Shariq Moore, Nick Nabar, Vick Nabar, Nikil Prasad, Joshua Speagle, Sai Vangala
Teacher: Louise Thompson
School: Brookfield Central High School, Brookfield, WI
Mentor: Andrea Ferrante, M.D., Blood Center of Wisconsin
Accumulation of excess iron results in a common hereditary disease, Hereditary Haemochromatosis (HH). There are various genetic mutations that lead to different forms of the disease. HH-I is a form of this disease in which iron accumulates in hepatocytes and intestinal epithelial cells and is associated with a mutation in the HFE (high iron protein) gene. The Brookfield Central SMART Team (Students Modeling A Research Topic) developed a model of HFE using 3D printing technology. The HFE gene encodes for a non-classical MHC class I protein. In physiological conditions, HFE is expressed and translocated to the cell surface where it may interact with a transferrin receptor (Tfr). The binding of the α1/β2 domains of HFE to the Tfr allows for controlled release of iron bound to transferrin-transferrin receptor complex. A mutation (845G>A) causes the replacement of a cysteine with a tyrosine (C282Y). This replacement prevents the α3 subunit of HFE from folding properly and from interacting with β2 microglobulin, abrogating the translocation of the HFE-microglobulin complex to the cell membrane and promoting its rapid degradation. This defect hinders the regulatory capability of HFE. Current treatments include phlebotomy to prevent organ damage from accumulated iron. Further study to increase understanding of the regulatory mechanism may lead to improved treatment design.
I'm a PC (Pyruvate Carboxylase) ... and diabetes was not my idea!
Brown Deer High School
Students: S. Andera-Cato, A. Arnold, S. Bach, J. DeSwarte, A. Faught, E. Frisch, A. Her, A. Keller, E. Kennedy, T. Martin, D. McMurray, C. Mitch, C. Orozco, C. Rice, B. Roberts, A. Rodgers, A. Sauer, A. Schulman, G. Sekhon, A. Suggs, K. Surfus, S. Tucker, T. Wray
Teacher: Gina Vogt
School: Brown Deer High School, Brown Deer, WI
Mentor: Martin St. Maurice, Ph.D., Marquette University
NIH estimates that 23 million Americans have diabetes, and 6.2 million are undiagnosed. If untreated, diabetes can cause complications, including heart disease and neuropathy. Type two diabetes patients cannot regulate glucose due to insulin resistance or deficiency. Pyruvate carboxylase (PC) plays an important role in insulin release from pancreatic β cells. Abnormal PC activity has been correlated with type two diabetes. PC is a dimer of dimers, each monomer a single chain with four domains: N-terminal biotin carboxylase (BC), central carboxyltransferase (CT), C-terminal biotin carboxyl carrier protein (BCCP), and allosteric domains. PC catalyzes the conversion of pyruvate to oxaloacetate (OAA). The process begins when biotin is carboxylated at the BC active site. The BCCP domain transfers the carboxybiotin to an active site in the CT domain. OAA is formed at the CT domain by adding a carboxyl group to pyruvate. Researchers concluded that the BCCP domain swings between active sites on opposite chains, instead of sites on the same chain. The Brown Deer SMART Team (Students Modeling A Research Topic), in collaboration with MSOE, built a model of PC using 3D printing technology illustrating this movement of the BCCP domain. Current research is focused on increasing PC activity through controlling a binding site in the allosteric domain, which may increase insulin production.
Isovaleryl-CoA Dehydrogenase: Dehydrate This!
Cedarburg High School
Students: Beth Bougie, Matt Cira, Colin Erovick, Anne Fahey, Nick Grabon, Kelsey Jeletz, Eleanore Kukla, Matt Murphy, Tim Rohman, Alyssa Sass, Michelle Sella, Laura Tiffany, Sam Wolff
Teacher: Karen Tiffany
School: Cedarburg High School, Cedarburg, WI
Mentor: Jung-Ja Kim, Ph.D., Medical College of Wisconsin
An OveRASsertive Mutation: The Uncontrolled Signaling in Cancer-Causing Ras
Grafton High School
Students: Katie Best, Lisa Borden, Kelsi Chesney, Emily Dufner, Elizabeth Fahey, Matt Harter, Alex Konop, Gabi Kosloske, Kaleigh Kozak, Michaela Liesenberg, Sadie Nennig, Nick Scherzer
Teacher: Dan Goetz
School: Grafton High School, Grafton, WI
Mentor: Casey O'Connor, Medical College of Wisconsin
Ras is a signaling protein that acts to control certain cell functions, such as cell division, gene expression, or cell-to-cell communication. Ras is located in the cytoplasm near the cell membrane and is activated by the exchange of GDP for GTP. Ras deactivation is accomplished by hydrolysis of GTP to GDP. A mutation in Ras that prevents hydrolysis of GTP is found in a staggering number of cancerous cells and results in uncontrolled signaling of Ras-regulated pathways. Understanding the multiple structures and dynamics of RasGTP may lead to treatments for cancers with a permanently active Ras. When Ras is activated, it binds to other signaling proteins called effectors, which determine the cell functions to be carried out. The effector RalGDS is activated when bound to RasGTP, and leads to exchange of RalGDP for RalGTP. A single amino acid mutation in Ras permanently traps the protein in an active state with GTP, leading to increased association with RalGDS, resulting in an exchange of RalGDP for RalGTP. When Ral is activated, excessive signaling for cell proliferation and anti-tumor suppression is favored along with a change in cell shape. A critical step for developing possible anti-Ras cancer treatments lies within a complete understanding of the interaction between RasGTP and its effectors like RalGDS.
Cabin1's Control of MEF2 in the Developing Central Nervous System
Greenfield High School
Students: Robert Bhatia, Ellen Campbell, Chelsea Herr, Leticia Gonzalez, Alohani Maya, Danielle Murphy, Tiann Nelson-Luck, Kara Stuiber, and Jordan Tian
Teachers: Julie Fangmann and Martin Volk
School: Greenfield High School, Greenfield, WI
Mentors: Dena Hammond and Ava Udvadia, Ph.D. University of Wisconsin - Milwaukee, WATER Institute
Cabin1 (calcineurin binding protein) is predicted to play an important role in maintaining the nervous system, which regulates important functions such as breathing, heart rate, thinking, and movement. Mice lacking Cabin1 die early in development, and other Cabin1 malfunctions have been linked to cancer. As the nervous system develops, neurons require guidance to determine their growth. The expression of specific proteins influences this neuronal growth. Transcription factors, proteins that bind to DNA and other proteins, regulate the production of these neuronal growth proteins. MEF2 (myocyte enhancer factor 2), a protein known to bind to Cabin1, is involved in nervous system development. MEF2 is a transcription factor necessary for neuronal growth and survival. MEF2 activates transcription when it binds to DNA, causing proteins involved in neural development to be made. MEF2 has a hydrophobic binding pocket that attracts four amino acids found on Cabin1: Ile106, Thre110, Ile116, and Leu119. When Cabin1, a transcription repressor found in the nucleus, binds to MEF2's hydrophobic binding pocket, transcription is turned off. This prevents the proteins from being produced and causes neurons to stop growing or to potentially die. Since cell death and survival are both necessary for nervous system development, Cabin1 is hypothesized to play a major role in this process. Current research is examining what happens to neuronal growth and survival when there is a shortage or an excess of Cabin1, eventually leading to a better understanding of the precise way Cabin1 functions.
"Like a Boss" The Role of FoxD3 in Pluripotency
Homestead High School
Students: Laura Auiar, Claudia D'Antoine, Sophia D'Antoine, Lisa Liu, Sarah Lopina, Sahitya Raja, Nikhil Ramnarayan, Daniel Schloegel, Ross Schloegel, Claire Songkakul, Rahul Subramanian, Bryanna Yeh
Teachers: Chris Schultz and Rick Hubbell
School: Homestead High School, Mequon, WI
Mentor: Sarah Kohler, Ph.D., Medical College of Wisconsin
Plexin: Putting Together the Pieces
Kettle Moraine High School
Students: Jacob Angst, Samantha Cinnick, Greg Dams, Allie Greene, Anna Henckel, Bronson Jastrow, Nick Merritt, Bradley Wilson, Aaron Zupan
Teacher: Steve Plum
School: Kettle Moraine High School, Wales, WI
Mentor: Shama Mirza, Ph.D., Medical College of Wisconsin
The cause of Proteus Syndrome, characterized by uncontrolled cell division leading to tumor formations, is currently not understood. Recent work suggests that cells from Proteus Syndrome patients express a higher level of a protein called Plexin D1. Plexin D1 has been found in angiogenic vessels during embryogenesis and may play an important role during embryonic development. Exploring the structure and function of Plexin D1 may contribute to a further understanding of Proteus Syndrome. Mass spectrometry can be used to compare the types and amounts of proteins found in diseased and healthy cells. Using Trypsin, the proteins in a healthy control cell and a diseased cell can be cleaved at basic residue groups along the polypeptide chains, such as arginine and lysine. Performing trypsin digestion in presence of heavy water (H218O) produces peptides with a +4 da mass shift. If 18O labeled peptides from diseased cells are mixed with unlabeled peptides from control cells, the ratio of labeled: unlabeled protein can indicate a change in protein expression between the two samples. The molecular weight of peptides from Proteus cells can be found through the process of electrospray ionization mass spectrometry (ESI-MS). This process involves a fine spray of protein fragments that can be analyzed by a computer. The data is then recorded in order to find the molecular weight of the peptides. Comparing the observed masses to a database of known proteins allows researchers to identify the peptide fragments. Plexin D1 may be the key to unlocking the mystery of Proteus Syndrome; the higher frequency of Plexin D1 in Proteus cells versus healthy cells remains promising. By collecting and analyzing data through mass spectrometry the uncertainty surrounding this elusive protein and obscure disease could be dissolved.
Cofacially-Arrayed Polybenzenoid Nano Structures
Marquette University High School
Students: Zeeshan Yacoob, Caleb Vogt, Patrick Jordan, Jose Rosas, Kienan Knight-Boehm, Hector Lopez, Fernando Buchanan-Nogueron, Nicholas Zausch, Qateeb Khan, Judson Bro, Payton Gill, Jacob Klusman, Alexander Lessila, Jed Sekaran, Caden Ulschmid
Teachers: Keith Klestinski and David Vogt
School: Marquette University High School, Milwaukee, WI
Mentor: Rajendra Rathore, Ph.D., Marquette University
Based on Moore's Law, every 18 months the number of transistors that can be placed per unit area on a microchip will double. However, by the year 2017 this trend will cease to exist due to the size restrictions imposed by silicon, the main component of microchips; this has led to the birth of molecular electronics. An organic molecule that has been extensively tested for its potential in molecular electronics is DNA. However, recent research has shown that DNA cannot function as a molecular wire due to its fragile nature. This is why Raj Rathore's group is developing organic molecular wires based on robust macromolecular structures. The specific assembly designed to study the wire behavior consists of a triad which is made up of polyfluorenes as an electron donor site, a spacer unit (or wire) made of polyphenylenes, and an electron donor-acceptor complexation site composed of hexamethylbenzene (HMB). A chloranil molecule - an electron acceptor - complexes with the HMB of the triad, and when a laser is shined on HMB/CA complex, a hole is introduced in the molecular wire which will travel 30 Å to the polyfluorene donor, via an electron hopping mechanism. The Marquette University High School SMART Team (Students Modeling A Research Topic) created a physical model of this molecular wire using 3D printing technology.
The Role of β-Catenin in Cell Division and Colon Cancer
Messmer High School
Students: Nancy Alba, Darrell Anderson, TeAngelo Cargille Jr., Sara Kujjo, Giovanni Rodriguez, Karla Romero
Teacher: Carol Johnson
School: Messmer High School, Milwaukee, WI
Mentors: Malathi Narayan and Sally Twining, Ph.D., Medical College of Wisconsin
Colon cancer is the fourth most lethal cancer in the U.S. As food passes through the colon, water and vitamins are absorbed and epithelial cells are sloughed off and replaced by a tightly regulated cell division process. Unregulated cell division can lead to the formation of polyps and tumors. β-catenin plays a role in regulating cell division and was modeled by the Messmer SMART Team (Students Modeling A Research Topic) using 3D printing technology. In non-dividing cells, a multi-protein complex of APC, GSK-3 and Axin phosphorylates β-catenin, signaling its degradation and preventing cell division. When cell division is needed, a Wnt signal cascade causes the complex to release β-catenin, stabilizing the protein for nuclear translocation and binding to TCF, a transcriptional activator, thus triggering cell division. Competitive binding of the inhibitor proteins, ICAT and Chibby, to β-catenin negatively regulates this process. In colon cancer, mutations in APC or Axin impede binding of the complex to b-catenin, preventing degradation, leading to increased nuclear localization, binding to TCF and deregulated cell division. Additionally, survivin, an anti-apoptotic protein that enables tumor cell survival, is upregulated. Understanding β-catenin's structure could help design drugs to promote binding of inhibitors to prevent the unregulated cell division of cancer.APC: The Key to Colon Cancer (The APC Protein and its role in controlling the cell cycle)
Nathan Hale High School
Students: Melissa Gall, Laenzio Garrett, Nick Goldner, Zach Koepp, Valerie Lamphear, Peter Nguyen, Vivek Patel, Stefan Pietrzak, Evan Rypel, Ajay Sreekanth, Kyle Tretow, Tyler Zajdel
Teachers: Sue Getzel and Anne Xiong
School: Nathan Hale High School, West Allis, WI
Mentor: William Berger, M.D., VA Medical Center
Colorectal cancer affects 1 in 18 Americans, and is linked to mutations in the Adenomatous Polyposis Coli (APC) gene. The rapid division of colonocytes is regulated by the Wingless Type (WnT) signaling pathway, mediated by β-catenin. In the nucleus, β-catenin binds to Transcription Cell Factor (TCF) and initiates transcription of cell cycle proteins. Alternatively, β-catenin binds to the 20-amino acid repeat region of the APC protein with the help of the scaffold protein axin. The enzyme GSK-3 then phosphorylates threonine and serine residues of the APC protein and subsequently β-catenin. Phosphorylated β-catenin is degraded, slowing mitosis. Mutations in APC allow β-catenin to accumulate, resulting in hyperproliferation of colonocytes, an early step in colon cancer development As such, better understanding of APC and its function could potentially lead to better diagnosis and treatment in colorectal cancer.
Lighting Up Science: Firefly Luciferase
St. Dominic's Middle School
Students: Elana Baltrusiatis, Allyson Bigelow, Rachel Brielmaier, Pamela Burbach, Jacob Dowler, John Fuller, Caroline Hildebrand, Teagan Jessup, Molly Jordan, Josh Kramer, Jenna Lieungh, Alex Mikhailov, Patrick O’Grady, Andrew Pelto, Quin Rowen, Rachelle Schmude, Robert Schultz, Katherine Seubert, Alex Sherman, Parker Sniatynski, Alex Venuti, Erin Verdeyen, Molly Wetzel
Teacher: Donna LaFlamme
School: St. Dominic Middle School, Brookfield, WI
Mentors: Nathan Duncan and Françoise Van den Bergh, Ph.D., Medical College of Wisconsin
Luciferase is the generic name for an enzyme responsible for bioluminescence reactions and is commonly associated with fireflies. It is also found in many other organisms including bacteria, fungi, anemones, and dinoflagellates. Since the gene for the North American firefly (Photinus pyralis) luciferase was cloned in 1985, scientists have been genetically engineering the gene into living cells. The luciferase reaction is now widely used in scientific research to study protein production in cells, to analyze gene promoter activity, to study stem cell function in vivo, and in cancer studies, to trace the metastasis of cancer cells in living test animals. The scientific study of the luciferase enzymes themselves is also continuing. In recent research, single amino acid mutations to the active site cause the emission of different colored light in a predictable way. The uses of and improvements in bioluminescent imaging are increasing exponentially in cell biology, molecular biology, and in medical research.
Go with the Flow: The Importance of VEGF-B
St. Joan Antida High School
Students: Beanca Buie, Maritza Campos, Brianna Castanon, Kateri Duncklee, Jagnoor Grewel, Monica Harris, Neli Jasso, Meghan Krause, Malayia Roper, Shinny Vang, Sukaina Yacoob
Teachers: Linda Krause and Cynthia McLinn
School: St. Joan Antida High School, Milwaukee, WI
Mentor: Mary Holtz, Ph.D., Medical College of Wisconsin
Blood vessels are such a vital part of out body that without them we would not progress from the embryo stage in the womb and our wounds would never heal. In addition, they are extremely helpful for the transportation of nutrients and oxygen and the removal of waste from cells in our body. In order for blood vessels to function correctly, Vascular Endothelial Growth Factor Type-B [VEGF-B] needs to be present. Unlike VEGF-A, which controls the development of blood vessels, VEGF-B controls the functions and prevents apoptosis. Though this protein does not make the blood vessels, VEGF-B stops apoptosis, or cell death, of endothelial cells, making it one of the main factors to the survival and development of blood vessels. VEGFs are a family of secreted glycoproteins, critical for development and function of blood vessels. The structure of VEGF-B consists of two cysteine knots, meaning two monomers, each of which contains eight cysteine residues and four disulfide bonds. The monomers are attached to form the dimer known as VEGF-B. VEGF-B promotes blood vessel survival by blocking apoptosis, in three kinds of cells that make up blood vessels: endothelial cells, smooth muscle cells, and pericytes. VEGF-B is sent between vascular endothelial cells (VEC) in the vessel connecting to the target cell in a locking fashion, assessing whether cells need to migrate, proliferate, specialize, or survive. Further understanding of the role that VEGF-B plays in vessel formation could be potentially used therapeutically against tumor formation. Tumors require angiogenesis, the production of new blood vessels, to enable blood transport to the tumor cells for continued growth. As a result, VEGF-B's abilities are taken advantage of by invading malignant tumors. Knowing more about VEGF-B is potentially life-saving and can be used to help maintain healthy blood vessels without encouraging vessel growth in tumors.
Inhibiting Dihydrofolate Reductase as a Treatment for Tuberculosis
Valders High School
Students: Corrine Brandl, Kaylin Kleinhans, Emily Weyker, Nicole Maala, Lauren Brandl, Grace Ebert, Paige Neumeyer, Ryan Jirschele, Gavin Schneider, Luke Schuh
Teacher: Joe Kinscher
School: Valders High School, Valders, WI
Mentors: Andrew Olson and Dan Sem, Ph.D., Marquette
University One-third of the world's population is infected by Mycobacterium tuberculosis (M.tb). Two million people die each year from tuberculosis (TB), the disease caused by this bacterium. TB primarily affects the lungs and is easily transmitted. One way to kill M. tb might be to inhibit the enzyme dihydrofolate reductase (DHFR). DHFR catalyzes the production of tetrahydrofolate by transferring a hydrogen ion (H-) from NADPH to dihydrofolate, thereby releasing tetrahydrofolate and NADP+. Tetrahydrofolate is essential to the bacteria's survival, and is a cofactor that is needed for the synthesis of the DNA base thymine. Isoniazid is one antibiotic already used to treat TB by targeting several TB proteins that are necessary for building M. tb cell walls and inhibiting DHFR. Unfortunately, strains of M. tb are evolving resistance to isoniazid, so next generation antibiotics are needed. A variation of isoniazid could be designed to avoid resistance, and to inhibit DHFR, thereby targeting bacterial DNA synthesis. The Valders SMART Team (Students Modeling A Research Topic), using 3D printing technology, created a physical model of DHFR and a possible inhibitor of tetrahydrofolate production.
EnvZ: Triggering Steinernema carpocapsae's Secret Weapon
Wauwatosa West High School
Students: Waj Ali, Jimmy Kralj, Jordan Llanas, Leah Rogers,
Mariah Rogers
Teacher: Mary Anne Haasch
School: Wauwatosa West High School, Wauwatosa, WI
Mentor: Steve Forst, Ph.D., University of Wisconsin - Milwaukee
The bacterium Xenorhabdus nematophilia participates in an unusual and fascinating mutualistic relationship with the nematode, Steinernema carpocapsae, which could not complete its lifecycle without the bacteria's help. EnvZ, a kinase protein located in the cell membrane of the bacterium, is critical to both organisms' success. Xenorhabdus resides quietly in a specialized pouch in Steinernema's intestines. To reproduce, the juvenile nematode enters an insect host via the anus or the mouth, and bores a hole though the intestine wall to get into the insect's blood. In response to the environmental signals in the blood, the nematode's pharynx begins to pump, forcing the Xenorhabdus out of the intestinal pouch and into the insect's blood. Xenorhabdus begins to act as an insect pathogen - killing the host insect, while simultaneously secreting antibiotics to eliminate its bacterial competitors. In killing the insect host, Xenorhabdus provides its symbiotic partner, Steinernema, with a carcass in which to reproduce. EnvZ helps Xenorhabdus sense the higher solute concentrations in the insect's blood through a currently unknown mechanism. EnvZ functions as a dimer. In response to appropriate environmental signals, a phosphate from ATP bound to the cytoplasm section of one EnvZ molecule is transferred to the HIS243 of another EnvZ. The phosphate group on HIS243 is then transferred to OmpR, a gene-regulating protein in Xenorhabdus that regulates genes that produce the antibiotics used to kill bacterial competitors. OmpR also regulates genes for exoenzymes that degrade insect tissues providing nutrients that help the nematode to reproduce.
The Dirt is Mightier than the Sword: Tetanus Toxin in the Human Body
Whitefish Bay High School
Students: Cara Ahrenhoerster, Quinn Beightol, Gabe Drury, Xavier Durawa, Riley Gaserowski, Shirley Hu, Domi Lauko, Ciara Otto, Andrew Phillips, Helen Wauck
Teachers: Paula Krukar and Michael Krack
School: Whitefish Bay High School, Whitefish Bay, WI
Mentor: Joseph T. Barbieri, Ph.D., Medical College of Wisconsin
Tetanus neurotoxin (TeNT), one of the most potent toxins known to humans, causes paralytic death to thousands of humans annually. TeNT is produced by the bacterium, Clostridium tetani, an anaerobic bacterium usually found as spores in soil. C. tetani often infects humans through open wounds where the bacterium colonizes the infected tissues. There are two domains of TeNT: the A domain possesses catalytic activity, while the B domain is made up of two sub-domains: the translocation sub-domain and receptor-binding sub-domain.
The SNARE protein contains VAMP-2, the target of the catalytic A domain of the TeNT. The SNARE protein regulates fusion of synaptic vesicles with the plasma membrane of the neuron, allowing the release of neurotransmitters that are responsible for relaying nerve signals such as inhibitory impulses to the body's muscle cells. TeNT binds to two gangliosides, on the presynaptic membrane of α-motor neurons. TeNT hijacks the trafficking machinery of the motor neuron and moves to the central nervous system, where it again binds to two gangliosides on the surface of neurons. Next, it enters the neuron by an endocytic mechanism to release the catalytic A domain into the host cell cytosol. The catalytic A domain cleaves the SNARE protein, which inhibits neurotransmitter fusion to the host cell membrane and the release of inhibitory neurotransmitter molecules. The loss of inhibitory impulses results in reflex irritability, autonomic hyperactivity, the classic large muscle spasticity and lockjaw associated with tetanus.