This website uses cookies to ensure you have the best experience. Learn more

Study Of New Biotinylated Analogue Of Dequalinium For Protein Kinase C??Inhibition And Identification Of The Binding Sites

2027 words - 8 pages

Specific Aims

Protein Kinase Cα (PKCα) is well known for its role in cellular signaling pathways that govern normal cell growth and differentiation. Due to its role in tumor formation and metastasis, PKC is being studied as a target for new chemotherapeutic agents. From 1987, Dequalinium (DECA) (quinolinium, 1,1’-(1,10-decanediyl)-bis(4-amino-2-methyl diiodie) has been studied as potent antitumor agent that recognizes several isoforms of PKC in vitro and in cells.

C10-DECA consists of two quinolinium moieties linked by a ten-carbon methylene bridge. Previous studies have shown that, PKC activity in vitro can be irreversibly inhibited when treated with 250nM of DECA and irradiated with 366nm of light. DECA directly interacts with catalytic domain of PKCα such that ring moieties bind at two distinct sites in a trans-oid fashion. The identity of these two sites remains unknown. Previous studies show that PKCαthe linker length of DECA. Studies have shown that maximal inhibition is achieved in vitro with a C-14 alkyl linker. In this project we are using biotinylated analogue of C14-DECA to study its PKC inhibition and identify the binding sites. We hypothesize that this DECA analogue will bind the catalytic sites of PKCα and thus inhibit its activation. Following binding, PKCα-DECA complex will be pulled down and further analyzed for the identification of binding sites.

The specific aims of this project are:

1. To identify the catalytic binding site
a. To isolate PKCα and biotinylated C14-DECA complex.
After photoinduced inactivation of PKCα with biotinylated C14-DECA, the complex will be isolated using streptavidin-agarose resin.
b. To identify the ligand bound peptide fragment.
The purified PKCαDECA complex will be trypsinized and the fragments will be identified by MALTI-TOF.

2. To do a comparative study of PKCα inhibition by new biotinylated analogue of C14-DECA vs conventional C14-DECA.
The biotinylated DECA analogue should have similar PKC inhibition compared to C14-DECA analogue. It will be studied using phosphotransferase activity of PKCα.

3. To determine cell motility inhibition by new biotinylated analogue of C14-DECA.
Since DECA is a potent PKCα inhibitor, inhibition of this enzyme should limit motility of cancerous cells. Wound healing assay will be performed to measure the motility of cells under influence of DECA drugs.

The long-term goal of this project is to develop better PKCα inhibitors. From our experiments we will determine if the biotinylated C14-DECA analogue is as much efficient as conventional C14-DECA analogue for PKCα inhibition. The biotin tag will assist in pulling down and isolating probed PKCα and thus utilize it for identification of binding sites. The identification of binding site will provide important information on further modification of DECA and development of new drugs for PKCα inhibition.

Background and Significance

PKCαand Cancer: ...

Find Another Essay On Study of New Biotinylated Analogue of Dequalinium for Protein Kinase C??Inhibition and Identification of the Binding Sites

Short-term in vitro Inhibition of Glycogen

1578 words - 6 pages -1 on tyrosine residues. Tyrosine phosphorylation activates IRS-1 allowing it to interact with and activate the next enzyme in the pathway, PI3-kinase, and continue the downstream process that brings glucose into the cell for glycogen synthesis3. Henriksen’s paper outlines the results of a study that investigated the inhibition of GSK-3 on skeletal muscle from overtly type 2 diabetic Zucker Diabetic Fatty (ZDF) rats. Inhibition of GSK-3

The Pros and Cons of Ethnic Identification

832 words - 3 pages minority works hard to spread the French language. Jews who have immigrated to Australia still practice their religious habits. Briefly, ethnic identification is necessary to people’s psychological well-being but we can’t deny that the idea of races is a problematic concept. Race and Ethnicity are controversial terms that are defined and used by people in many different ways. Some persons say that an identity can’t be fixed that’s why a study

The Identification and Treatment of OCD

1305 words - 5 pages possible risk factors are family history of the disorder and traumatic experiences. For instance, in the case study of a woman named Sophie who was diagnosed with OCD, there were many conceivable contributing factors. Her parents had “mental health” issues and she was bullied at school, and then after being told that she could not give birth, she gave birth to three children. With regard to the pregnancy and birth of her youngest (the result of an

In-silico analysis of phytocompounds for inhibition of nAChRs receptor mediated lung cancer

2034 words - 8 pages antitumour activity against a broad range of rodent tumors (Chia-Ping Huang Yang, et al., 2000; Kandaswami C, et al 2005). The major cellular target for Taxol is the tubulin / microtubule system of the cancer cell. Taxol has a specific binding site on the microtubule and incubation of cells with Taxol causes the formation of stable bundles of microtubules that disrupt the normal polymerization/depolymerization cycle of microtubules and suppresses

Is Whey Protein the way to go?-Research Paper on the benefits and drawbacks of using whey protein for weight training

3045 words - 12 pages 360) which helps with healing the muscle after working out. Branched-chain amino acids are special amino acids because they are taken from the blood and broken down by the cells into glucose for energy. (Pasin 2000)Methods of ManufacturingIon-ExchangeWhey protein can be manufactured threw a process known as ion-exchange. Here the "protein molecules are removed from liquid whey threw chemical binding to specially developed resins, across which the

Impact of the Presence of Protein Aggregates

1822 words - 8 pages working with pre-selected, well-characterized proteins, especially at the early development and formulation stages. This emphasizes the need for new in silico tools than can be used to rank protein-based drugs, and help in the selection of the proper molecules for product development. Despite the above-described algorithms perform fairly well in detecting aggregation-nucleating sequences in proteins primary sequences it is also true that in the

Characterizing the mRNA-binding activity of MEX-5

1747 words - 7 pages . Because for CCCH-type TZF proteins the architecture of the backbone contributes to the mechanism of specific RNA sequence recognition, the knowledge of the structure of the TZF domain of MEX-5 will provide insight in the pattern of interactions that defines the RNA-binding specificity. Strategy: In order to avoid issues caused by the instability of the protein in the unbound state, the structure of MEX-5268-346 will be obtained as a complex with a

Identification of the Dairy Propionibacteria

541 words - 3 pages for strain identification and genotyping (Gautier, Carvahalho, Rouault, 1996; Rossi, Torriani, Dellaglio,1998). As we showed previously in Blasco et al 2011, the ability to metabolise aspartate is depending on the characteristics of strains and it is a very important factor to consider when selecting new cultures. Even though excessive aspartase activity has hidden dangers such as late fermentation, a moderate activity may influence the

The Journey of Self-Identification

1443 words - 6 pages According to behavioral psychologists, at birth children begin to identify themselves through parental figures of the same sex, “especially to those the child depends on for love and care.” The process typically encourages children to imitate their parental figures, and therefore limits the growth of identification through one’s self. In the novel Their Eyes Were Watching God by Zora Neale Hurston, the character Janie is used to show the

The Identification Technology of Biometrics

701 words - 3 pages The recent advances of information technologies and the increasing requirements for security proposed have led to a rapid development of automatic personal identification systems based on biometrics. Biometrics refers to identify an individual based on his or her distinctive physiological (e.g., fingerprints, face, retina, iris) or behavioral (e.g., gait, signature) characteristics accurately [1]. It is inherently more reliable and more capable

Inhibition of Nitric Oxide Synthase Impairs a Distinct Form of Long-Term Memory in the Honeybee

3416 words - 14 pages inhibition of nitric oxide synthase only effects long term memory obtained in multiple conditioning trials. The difference in response can be seen only when the bees' memories were tested 24 hours after treatment. Impact Every day we experience new and also routine occurrences. There was a day that we smelled and tasted an orange for the first time. Since that time, we have smelled and eaten oranges enough to know that when a specific "orange

Similar Essays

Runx2 Binding Protein And The Regulation Of Osteogenesis

1964 words - 8 pages (Wang et al., 2013). Tumor suppressing subtransferable candidate 1, TSSC1, is a target of the binding protein known to inhibit breast cancer cell invasion (Wang et al., 2013). Because Runx2 negatively regulates TSSC1, overexpression of the binding protein results in bone osteolysis (Wang et al., 2013). By studying the TSSC1-dependent nature of this cancer, results could be used for treatments of breast cancer and to prevent metastasis (Wang et

The Country Club College Inhibition Of Eudaimonia

1985 words - 8 pages based on pleasure. Removing consumption spending from the pinnacle of college spending can lead to colleges competition based on academics and the utilization of reason. As a result, America can achieve a more educated populace and a mentality based on virtue. Works Cited “Consumption Amenities in Higher Education.” The Brookings Institution. N. p., n.d. Web. 8 Apr. 2014. Dillon, Sam. “Share of College Spending for Recreation Is Rising.” The New York Times 9 July 2010. NYTimes.com. Web. 8 Apr. 2014. “Many Students Opt for Colleges That Spend More on Nonacademic Functions, Study Finds | Inside Higher Ed.” N. p., n.d. Web. 8 Apr. 2014.

Commercial Transactions And The Elements Of A Legal Binding Contract

1114 words - 4 pages There are four requirements which constitute what is generally referred to as the elements of a legally binding contract. If only one of the four elements is not present, a legal bond is not able to be created. Listed below are the four elements of a valid contract;(1) Agreement: An agreement to shape a contract requires that it contain both an offer and an acceptance. This means that one party must proposition the entrance into a lawful

Regulation And Functions Of The P53 Protein

1189 words - 5 pages (PRR) - This plays a role in p53 stability 3. Central DNA binding domain (DBD) - required for binding to specific sites on the DNA. 4. Tetramerization domain (OD) - required for the assembly of the functional tetramer. 5. Carboxyl terminus domain (CTD) - which is bound to DNA binding domain and is involved in negative modulation of DNA binding domain. The central DNA binding domain is the most highly conserved region of p53, when compared to its