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Chemistry 2008
Summer Science Research Abstract
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Synthesis and Electrochemical Analysis of Transition Metal Complexes
with Various Functionalities for Surface Modification
Suzanne Ali
Mentor: Prof. Jonas Goldsmith
It is possible to functionalize various surfaces by attaching various different types of organic ligands to them. The organic ligands are able to form complexes with transition metals, and an electrode composed of the surface material in question is placed into a solution containing a low concentration of the aforementioned metal complex. The interactions of various ligands with metal and carbon surfaces can be analyzed electrochemically, and in this manner the rate at which these ligands adhere to the surface, based on varying concentration, can be examined.
During this summer research period, we will synthesize 4-methyl-4-(5-bromobutyl)-2,2’-bipyridine and 4-methyl-4-(5-bromononyl)-2,2’-bipyridine. The alkyl halide will be replaced with an amine group (see Fig. 1), and then an amide group (see Fig. 1) attached to a polyaromatic moity, such as pyrene, as seen in figure 3. The purity of each synthesized compound will be determined through Nuclear Magnetic Resonance and Mass Spectrometry. Due to the nature of the experiments, it is necessary that each step of the synthesis produce a particularly pure product. We will then complex this molecule with a transition metal, such as cobalt or ruthenium, as seen in figure 3. Due to the chelate effect, the preference for the formation of a transition metal complex with a bidentate ligand, the bipyridine molecule will form particularly strong complexes. The extended pi electron system which the polyaromatic group possesses will cause it to adsorb strongly to a graphite surface, which has a similar extended pi electron system. Due to this affinity for adsorption, we can use a graphite electrode to observe this process.
We will also examine, electrochemically, the interactions of 4-methyl-4-(5-thiobutyl)-2,2’-bipyridine and 4-methyl-4-(5-thiononyl)-2,2’-bipyridine with gold surfaces. The thiol group (see Fig. 1), which, as with the amine group, replaces the alkyl halide, has strong affinity for gold surfaces. This affinity will cause the thiol group to adsorb strongly to the surface of the gold electrode, and we will be able to examine the interactions of transition metal complexes containing this ligand, as shown in figure 2, with a gold electrode.
 
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Syntheses of [n]Phenacenes
Kate Butler
and
Amber Hopkins
Mentor: Dr. Frank Mallory
[n]Phenacenes are compounds that contain n benzene rings fused together in a zigzag pattern. An example of a [7]phenacene derivative is shown below.

R=alkyl group
The synthesis of [n]phenacenes is of particular importance for the investigation of whether pseudo one-dimensional versions of the pseudo two-dimensional graphite sheets possess similar patterns of conductivity to the graphite sheets. Previously, an [n]phenacene having 11 fused rings has been synthesized by the Mallory group. [n]Phenacenes with n > 6 are extremely insoluble, so to produce larger [n]phenacenes by chemical synthesis, solubilizing groups (R) must be attached. [n]Phenacenes with n > 11 have been attempted, but have proved unsuccessful due to issues of solubility based on the various R-groups used.
The most recently concluded study tested a branched 12-carbon chain and a straight 12-carbon chain as solubilizing groups, both of which proved to make the phenacene product too soluble. Currently, solubilizing groups of shorter carbon chains are the focus of our investigation in hopes of obtaining correct solubility. Two possible schemes contain 8-carbons each, one straight chain and one branched chain.
Using synthesis schemes beginning from 1-bromobutane and 1-bromooctane, a sequence of reactions will be explored in the attempted syntheses of [7]phenacene and [15]phenacene derivatives.
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Using Polymerase Chain Reaction-Restriction Length Fragment Polymorphism (PCR-RFLP)
method to establish Genetic Variation in Local Mushrooms and Flies
Clarah Chelagat Lelei
Mentor: Dr. Susan White
Restriction Fragment Length Polymorphism (RFLP) is a variation in the DNA sequence in a genome. In PCR-RFLP technique restriction enzymes plays an important role in DNA cleavage. Restriction enzymes recognize a particular short segment of double stranded DNA that consists of 4 to 12 base pairs. These sites are called restriction sites and they vary in different species because it can occur in the middle of a DNA strand or at the ends. The sites are also palindromic- that is the 5’ to 3’ sequence on one strand is identical to the 5’ to 3’ sequence on the antiparallel complementary strand. The segments resulting from restriction digest vary in size and thus can be detected using gel electrophoresis. For example, HinfI restriction enzyme recognition site is 5’TCGA…3’, HaeIII recognizes 5’GGCC…3’ and Taqα1 recognizes 5’…GANTC…3’ The length of the cleaved DNA varies, and the variation can be detected by performing a gel electrophoresis. The aim of the research is to find out if PCR-RFLP can be used to explain the diversity in the local mushrooms and flies based on the DNA sequence.
My summer research will involve extraction of DNA from local mushroom species and flies. I will then amplify the DNA I have extracted using the Polymerase chain reaction. Specific regions of the DNA will be amplified by thermostat Taq polymerase. A pair of single stranded DNA will act as replication primers and delimits the region of the target molecule that will be amplified. The amplification will be followed by a restriction digest using different restriction enzymes. Then the agarose gel electrophoresis will be performed to visualize the differences in the resulting restriction digest.
If the PCR-RFLP method works, the results from this experiment will help in exploring the genetic variation in mushrooms. The results might also help in explaining why some mushrooms are edible and why some are poisonous and why some insects harmful. The method might also be applied to other species and this may help in answering questions concerning molecular diversity.
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Synthesis and Characterization of DNA-Intercalating and Potential Photocleaving Ru(II)-bis(bipyridine)-Pteridinyl Complexes and Similar Co(III) Complexes
Erika Lippoldt
Mentors: Professor Sharon Burgmayer and Shannon Dalton
Abstract. Small transition metal molecules have been the object of many studies due to their interactions with DNA and the resulting effects on the regulation of DNA transcription and replication, as well as their potential as pharmaceuticals. Ru(II) compounds are especially useful as probes due to their stability and photophysical properties. Bis(bipyridyl) Ru(II) complexes of pteridinyl-phenanthroline ligands have been of particular interest because pteridinyl ligands possess H-bonding patterns complementary to the purine and pyrimidine bases of DNA and RNA. The pteridinyl ligand of these Ru-pteridine complexes is capable of inserting itself between the base pairs of DNA, thus binding to DNA via intercalation. Other metal complexes, including certain Co(III) complexes, have been known to cleave DNA in the presence of light (photoactivated cleavage of DNA). Such metallointercalators are practical for their high affinity for double-stranded DNA and because they include a range of redox-active metal centers and ligands.
In my summer research, I will synthesize five Ru-pteridinyl complexes previously studied in this laboratory by varying the pteridinyl-phenanthroline ligands, in addition to which I will synthesize the DNA intercalator [Ru(bpy)2(dppz)]2+, which has been much studied in other laboratories and is used here as a positive control. The multiple-step synthesis is similar for each complex: 1,10-phenanthroline-5,6-dione is synthesized and then reacted with a diamino-pyrimidine to obtain the various ligands, which are then coordinated to RuII by reacting them with Ru(bpy)2Cl2 to obtain the final Ru complex of the form [Ru((bpy)2(L)]2+, where L stands for any of the five ligands. The ligands of interest are L-amino, L-diamino, L-pterin, L-allox and L-Me2allox. All synthesis products are characterized using 1H-NMR, IR and ESI-MS. In addition to these syntheses, I will investigate the synthesis of analogous Co(III)-pteridinyl or other metal-pteridinyl complexes, which will be explored as DNA cleaving agents. DNA cleavage studies could lead to the determination of the metallointercalator-DNA binding sites as well as the discovery of useful DNA manipulation techniques and potential pharmaceuticals.
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Functionalizing Surfaces with Transition Metal Complexes
Erica Lo
Mentor: Dr. Goldsmith
The nanotechnology field is growing rapidly thus novel methods in modulating behavior on the nanoscale is necessary for the development of nanoelectronics. In order to control the behavior of this technology, one must appeal to redox chemistry. Adding functionality to surfaces is possible through the synthesis of metal complexes with the appropriate substituents, allowing adsorption to various surfaces.
The focus of this research is to achieve the synthesis of a transition metal complex capable of the aforementioned functionalization. First, a bromoalkyl chain of varying lengths is to be added to a 4,4’-dimethyl-2,2’-bipyridine molecule giving 4-bromobutyl-4-methyl-2,2’-bipyridine or 4-bromononyl-4-methyl-2,2’-bipyridine. The added bromine will then be replaced with a thiol, an SH group, which has the capability of adsorbing to gold surfaces. The bipyridine portion of the molecule also enables the formation of a stable metal complex. In this case the transition metal is ruthenium as shown in Figure 1.
The second goal is to synthesize a molecule with the capability of adsorbing to varying carbon surfaces. The first step is the same as the formation of the previous molecule. Once the molecule has a bromoalkyl substituent, the bromine will be converted to an amine. This NH2 group will allow the coupling of polyaromatic groups resulting in a ligand with an extended pi system. The ligands will then form a complex with a transition metal giving a product as seen in Figure 2 where the polyaromatic group shown is pyrene. The manner of how these molecules adsorb to gold, platinum surfaces, or carbon surfaces will then be observed by using electrochemistry.
 
Figure 1. Figure 2.
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Functionalization of a Variety of Carbon Surfaces with Transition Metal Complexes: Glassy Carbon, HOPG and SWNT
Eden McQueen
Mentor: Dr. Jonas Goldsmith
The behavior of several carbon surface types functionalized with the compound below (1) will be explored.

A sample of complex 1 was synthesized and re-crystallized to obtain a workable powder. Polished glassy carbon, highly oriented pyrolytic graphite, and single-walled carbon nanotubes (SWNT), were functionalized using 1 by dissolving the complex in acetonitrile and exposing the carbon surface to a dilute solution of 1. The HOPG and glassy carbon functionalized surfaces were characterized by the observed redox behavior on an electrode surface using cyclic voltammetry. Experiments used a range of concentrations from 0.1μM to 2.0μM, and were conducted over the period of 1.5 hours. For each data point the reduction peak height was recorded. The equation was used to obtain the electrode coverage for each data point. The kinetics and thermodynamics of the adsorption process can be determined by an analysis of coverage (Γ) versus time and coverage versus concentration. In order to study the adsorption behavior of 1 on SWNT, an additional step was required to disperse and separate the SWNT material. Suspensions of SWNT were created using several solvents, including Milli-Q water, chloroform, ethanol, and isopropanol. A range of surfactants, SDS, PVP, PVA, and Triton X-100, were also used and compared as to their relative utilities for suspension formation. Suspensions that appeared to be well-distributed were diluted and filtered. Atomic force microscopy was used to characterize the appearance of the SWNT suspensions. The dispersions will also be cast onto metal electrodes to study their adsorption behavior.
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A Thermal Denaturation method for distinguishing
between K-turn and non-K-turn RNA
Amber R. Moore
Mentor: Dr. Susan A. White
 
Abstract. Protein-RNA interactions are important for cellular growth and regulation. Proper interactions between protein and RNA require both of their interfacial sites to be conserved for specific recognition and binding. Organisms, like Saccharomyces cerevisiae (yeast), have evolved in ways that condense multi-component and multi-step pathways into more self-contained processes. This allows the cell machinery to maintain, if not improve, its specificity and regulation, while increasing the efficiency of these vital mechanisms. Knowledge of protein and RNA interactions continues to aid in the development of therapeutic approaches for the treatment of various diseases.
 The S. cerevisiae autoregulatory ribosomal protein L30e, or RPL30, is capable of inhibiting the splicing and translation of its own transcript and mRNA by forming a protein-RNA complex. The L30e protein has a single domain composed of four beta sheets and four alpha helices. The L30e RNA is present in two strands and assumes a helix-internal loop-helix motif, commonly referred to as the kink-turn, or K-turn motif. The internal loop, comprised of one strand with unpaired nucleotides and a second strand with missing nucleotides, connects a canonical stem (Watson-Crick base paired helix) to a non-canonical stem (helix containing some non-Watson-Crick base pairs). L30e recognizes and binds to the K-turn RNA when there is an excess of L30e protein (not being incorporated into the ribosome). This forms the L30e protein-RNA complex that represses further protein expression. We expect to find that any RNA sequence mutation altering the K-turn will disturb the protein-RNA interaction of the L30e complex.
Thermal denaturation will be the applied method in differentiating between K-turn RNA and non-K-turn RNA while Mg2+ is present to simulate cellular conditions and assist the K-turn RNA in maintaining its sharp bent structure. Results are expected to show differences in the stabilities among the RNA variants being tested: BP, KT12, and KTAU. The BP (base paired) variant has complete Watson-Crick base pairing among the nucleotides, whereas the KT (kink turn) variant has non-Watson-Crick base pairing which causes the helix-internal loop-helix motif in the RNA. KT12 has an adenine (A) replaced with a uracil (U) at the twelfth nucleotide position in the K-turn region of the RNA. KTAU has the same A to U mutation, but it occurs outside of the K-turn region. Thermal denaturation is not only expected to show differences in stability between the BP and KT RNA, but also between the KT variants with nucleotide mutations inside and outside of the K-turn region.
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Modeling the Molybdenum Cofactor
Rebecca Rothstein
Mentors: Dr. Sharon J. Nieter Burgmayer and Graduate Student Kelly Ginnion
The second row transition metal molybdenum, atomic number 42, is a naturally occurring element in a countless number of biological reactions. Assuming the role as a biological catalytic center, molybdenum occupies the catalytic site in more than forty essential enzymes, which control the oxidation-reduction reactions for a broad range of inorganic and organic substrates. Considering the ubiquitous nature of molybdenum enzymes, with its presence in everything from simple bacteria to human beings, it is no wonder that the element has a profound evolutionary history, contributing significantly to the biological function of virtually every living organism. From the human perspective alone, three molybdoenzymes—sulfite oxidase, xanthine oxidase and aldehyde oxidase—are essential for proper health.
Contrary to popular scientific belief, molybdenum has not survived the perils of evolution single-handedly but rather it is now believed that such enzymes are the chemical descendants of tungsten enzymes, whose origin was traced to the most ancient of organisms, archaebacteria. Consequently, it has been found that the structures and functions of tungsten enzymes in bacteria are similar to those of the molybdenum enzymes typically found in more complex species like human beings. Molybdenum and tungsten enzymes are the only biological molecules to utilize the dithiolene as a metal ligand. Together, the dithiolene ligand bound to the molybdenum center in all molybdenum enzymes has been identified by the name of the molybdenum cofactor (colloquially referred to as Moco) illustrated in figure 1 below.

Figure 1. Moco consists of a pterin structure (blue) fused to a pyran (red) ring bearing an exocyclic dithiolene group (green) that is the site of metal coordination.
The identity of the molybdenum cofactor, the catalytic site in pyranopterin molybdenum enzymes, was sought through studies of the molybdenum enzymes and through synthetic efforts to assemble compounds that reproduce key spectroscopic, structural, and reactivity properties of the catalytic Mo center. The Burgmayer laboratory has developed novel models for the molybdenum cofactor which incorporate key structural features in all Mo and W enzymes including a dithiolene chelate joined to a pterin. The goal of this summer laboratory experience will be to synthesize the precursors of Moco models and then to utilize the starting material in order to build the pterin-dithiolene ligand. Generally, the synthetic scheme will entail coupling a Mo-tetrasulfide complex with a pterinyl alkyne to produce [Tp*Mo(O/S)(pterin-dithiolene)] complexes in both Mo (4+) and Mo (5+) oxidation states. While previous work has focused on the characterization on the purified Mo (5+) species, there is a strong hope of exploring the fascinating redox chemistry associated with Mo (4+) in the near future
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Using Analytical Centrifugation to Identify and Study Kink-turned RNA
Ashton Shaffer
Mentor: Dr. Susan White
RNA (ribonucleic acid) makes it possible for the genetic instructions encoded in DNA to be used to assemble essential molecules like proteins. RNA’s structure allows it to participate in reactions like transcription, which is basically the production of a complementary copy of DNA, and splicing, which is the processing and editing of RNA prior to use for protein production. RNA’s secondary structure, when it is double stranded, tends to consist of nucleotides that base pair in the traditional Watson-Crick manner, where hydrogen bonds are formed only between guanine-cytosine and adenine-uracil, and form a helical configuration. It can, however, have a kink-turn, which contains three unpaired nucleotides followed by several nucleotides that do not exhibit Watson-Crick base pairing. L30, a protein found in yeast, has been found to autoregulate its transcription and splicing by binding to its own RNA transcript, which forms a kink-turn. Learning more about this mechanism of identification, binding, and regulation will provide insight into RNA-protein interactions, which makes kink-turned RNA and L30 particularly interesting.
Kink-turn RNA has previously been studied using radioactive isotopes in gel mobility experiments where the differences are only detected in the presence of Magnesium (Mg2+).
Our objective is to identify RNA with kink-turns without using techniques like radioactive gel electrophoresis and instead utilizing a process called analytical centrifugation. This alternate approach is based on the biophysical principles of hydrodynamics and observes the rate of migration of RNA as it is spun at high speeds in a fluid environment in order to infer its structure. We plan to test it both with and without Mg2+ in solution with the RNA to determine whether or not there are any discernable differences in the resulting RNA mobility. It has the added advantage of permitting us to study RNA in an environment that closely resembles what is actually present in a cell. Our hope is that analytical centrifugation will enable us to not only distinguish the difference between kink-turned and base paired RNA, but also to tell to what degree the RNA is kink-turned (larger and smaller angles). We are also interested to observe whether or not Mg2+ must be present in order to make these distinctions. Being able to differentiate kink-turned and base-paired RNA adds to our general understanding of how structural changes affect the ability of proteins and RNA to interact and the chemical mechanisms of transcription, splicing, and regulation.
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Exploration of the Conjugate Addition Reactions of Carbon Nucleophiles
to Birch Cope Sequence Products
Sarah Tabi
Mentor Dr. Malachowski
Generally, I will be experimenting with different carbon nucleophiles of Birch Cope sequence products for multiple conjugate addition reactions. Below [Scheme 1] includes the outline of the starting materials that I have been synthesizing in order to do this. The last three steps of this scheme illustrate the Birch Cope sequence that will produce the starting material necessary for the aforementioned reactions.

I will delve into other carbon nucleophiles as well, including organocuprates, cyanide, and ketone/ester/nitro enolates. In this process, the steric limitations of the nucleophiles will be revealed.
In conducting these reactions, the stereoselectivity of the conjugate addition will become evident. The progress in this will aid in the asymmetric synthesis of cyclohexyl rings that possess multiple tertiary and quaternary asymmetric molecular sites. These types of structures are found in numerous biologically active molecular structures, such as saudin, anisatin, and jiadefenin. This type of understanding will help in future research involving the synthesis of complex, biologically active natural products.
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Design Dithiocarbamate Inhibitors with Increase Binding Affinity
to Indoleamine 2,3-Dioxygenase (IDO)
Tu-Anh Vu
Mentor: Judith LaLonde
Indoleamine 2,3-dioxygenase (IDO) is an immunosuppressant enzyme that is the rate-limiting enzyme in the production of quinolinic acid from tryptophan through the kynurenine pathway. Researchers have suggested that tumors can elevate the levels of IDO and use it to facilitate an immune escape. Previous studies have shown that knockout mice without IDO perform better in chemotherapy treatments than mice with IDO. Thus, designing IDO inhibitors may improve patients' responses to caner treatments. In this study, we will design dithiocarbamate inhibitors with increased binding affinity to IDO. In previously published work, our group has shown that hydroxyl and thiol substitution on the benzyl ring of 4-phenyl-imidazole improve binding affinity. Docking calculations of the dithiocarbamate compounds predict that the dithiocarbamate group binds to the heme iron, while the amino-linked benzyl ring of the dithiocarbamate is buried in the back of the cavity. We will apply lessons learned from the -OH and -SH substitutions of phenyl-imidazole to the dithiocarbamate class of compounds. Computational chemistry methods will be used to vet in silico prototype compounds prior to synthesis. The genetic docking algorithm, Gold, is being used to evaluate twenty various substitutions of the amino-linked benzyl ring of the dithiocarbamate. Computational results will determine the best candidates for synthesis.
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