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The NCSU Biochemistry blog is a team effort by the faculty and students of the biochemistry department at NC State University to describe past and current research, science topics of general interest, our opinions of current events in science, and situations that we find humorous at a public university.
Clay Clark
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by Clay Clark in Biochem Blogs
What if I told you there may be a new way of determining whether non-cancerous cells have the potential to form malignancies. If this were possible, it may lead to earlier diagnosis and treatment. The method may sound like … Continue reading →... Read more »
Kobayashi, Y., Absher, D., Gulzar, Z., Young, S., McKenney, J., Peehl, D., Brooks, J., Myers, R., & Sherlock, G. (2011) DNA methylation profiling reveals novel biomarkers and important roles for DNA methyltransferases in prostate cancer. Genome Research, 21(7), 1017-1027. DOI: 10.1101/gr.119487.110
Yi, J., Dhir, M., Guzzetta, A., Iacobuzio-Donahue, C., Heo, K., Yang, K., Suzuki, H., Toyota, M., Kim, H., & Ahuja, N. (2012) DNA methylation biomarker candidates for early detection of colon cancer. Tumor Biology, 33(2), 363-372. DOI: 10.1007/s13277-011-0302-2
Marta Kulis, Manel Esteller. (2010) DNA methylation and cancer. Advances in Genetics, 27-56. DOI: 10.1016/B978-0-12-380866-0.60002-2
by Clay Clark in Biochem Blogs
A new paper published in PLoS Biology characterizes two bacterial death pathways Programmed cell death (PCD) in eukaryotes is a well-studied process that is used by organisms to maintain homeostasis. The mechanisms of PCD are under intense study because altered … Continue reading →... Read more »
Erental, A., Sharon, I., & Engelberg-Kulka, H. (2012) Two Programmed Cell Death Systems in Escherichia coli: An Apoptotic-Like Death Is Inhibited by the mazEF-Mediated Death Pathway. PLoS Biology, 10(3). DOI: 10.1371/journal.pbio.1001281
Vesper, O., Amitai, S., Belitsky, M., Byrgazov, K., Kaberdina, A., Engelberg-Kulka, H., & Moll, I. (2011) Selective Translation of Leaderless mRNAs by Specialized Ribosomes Generated by MazF in Escherichia coli. Cell, 147(1), 147-157. DOI: 10.1016/j.cell.2011.07.047
Belitsky, M., Avshalom, H., Erental, A., Yelin, I., Kumar, S., London, N., Sperber, M., Schueler-Furman, O., & Engelberg-Kulka, H. (2011) The Escherichia coli Extracellular Death Factor EDF Induces the Endoribonucleolytic Activities of the Toxins MazF and ChpBK. Molecular Cell, 41(6), 625-635. DOI: 10.1016/j.molcel.2011.02.023
by Clay Clark in Biochem Blogs
sciseekclaimtoken-4f580c3179515 Rapid bioorthogonal labeling of proteins By Clay Clark, @biochemprof There are a number of current methods for labeling proteins for imaging either in vitro and/or in live cells and organisms, including fusions with fluorescent proteins, dyes, tags (such as SNAP, … Continue reading →... Read more »
Lang, K., Davis, L., Torres-Kolbus, J., Chou, C., Deiters, A., & Chin, J. (2012) Genetically encoded norbornene directs site-specific cellular protein labelling via a rapid bioorthogonal reaction. Nature Chemistry. DOI: 10.1038/NCHEM.1250
Chen, I., & Ting, A. (2005) Site-specific labeling of proteins with small molecules in live cells. Current Opinion in Biotechnology, 16(1), 35-40. DOI: 10.1016/j.copbio.2004.12.003
Best, M. (2009) Click Chemistry and Bioorthogonal Reactions: Unprecedented Selectivity in the Labeling of Biological Molecules. Biochemistry, 48(28), 6571-6584. DOI: 10.1021/bi9007726
by Clay Clark in Biochem Blogs
Let there be { if (RhoGTP + Effector == ProteinComplex) { Luc1 + Luc2 = ActiveLuciferase; LIGHT = 1; } else if (RhoGTP + GAP == RhoGDP) { RhoGDP + Effector = NoProteinComplex; LIGHT = 0; } else if (RhoGDP … Continue reading →... Read more »
Anderson, E., & Hamann, M. (2012) Detection of Rho GEF and GAP activity through a sensitive split luciferase assay system. Biochemical Journal, 441(3), 869-879. DOI: 10.1042/BJ20111111
Jaiswal M, Dubey BN, Koessmeier KT, Gremer L, & Ahmadian MR. (2012) Biochemical assays to characterize Rho GTPases. Methods in molecular biology (Clifton, N.J.), 37-58. PMID: 22144266
Massoud, T., Paulmurugan, R., De, A., Ray, P., & Gambhir, S. (2007) Reporter gene imaging of protein–protein interactions in living subjects. Current Opinion in Biotechnology, 18(1), 31-37. DOI: 10.1016/j.copbio.2007.01.007
by Clay Clark in Biochem Blogs
Recently I posted a blog on using miRNA profiling as biomarkers for cancer. Protein profiling is another potential tool for hunting biomarkers. Traditional microarray data are based on the assumption that each individual protein contributes independently to clinical outcomes. … Continue reading →... Read more »
Chuang, H., Lee, E., Liu, Y., Lee, D., & Ideker, T. (2007) Network-based classification of breast cancer metastasis. Molecular Systems Biology. DOI: 10.1038/msb4100180
Li, C., & Li, H. (2008) Network-constrained regularization and variable selection for analysis of genomic data. Bioinformatics, 24(9), 1175-1182. DOI: 10.1093/bioinformatics/btn081
Chen, L., Xuan, J., Riggins, R., Clarke, R., & Wang, Y. (2011) Identifying cancer biomarkers by network-constrained support vector machines. BMC Systems Biology, 5(1), 161. DOI: 10.1186/1752-0509-5-161
by Clay Clark in Biochem Blogs
No one wants to catch the flu! At the very least, it will put you out of commission for a week, and it can also cause life-threatening infections – pneumonia is the most common, although other bacterial diseases like … Continue reading →... Read more »
Monto, A., McKimm-Breschkin, J., Macken, C., Hampson, A., Hay, A., Klimov, A., Tashiro, M., Webster, R., Aymard, M., Hayden, F.... (2006) Detection of Influenza Viruses Resistant to Neuraminidase Inhibitors in Global Surveillance during the First 3 Years of Their Use. Antimicrobial Agents and Chemotherapy, 50(7), 2395-2402. DOI: 10.1128/AAC.01339-05
Yen, H., Herlocher, L., Hoffmann, E., Matrosovich, M., Monto, A., Webster, R., & Govorkova, E. (2005) Neuraminidase Inhibitor-Resistant Influenza Viruses May Differ Substantially in Fitness and Transmissibility. Antimicrobial Agents and Chemotherapy, 49(10), 4075-4084. DOI: 10.1128/AAC.49.10.4075-4084.2005
Hurt, A., Holien, J., & Barr, I. (2009) In Vitro Generation of Neuraminidase Inhibitor Resistance in A(H5N1) Influenza Viruses. Antimicrobial Agents and Chemotherapy, 53(10), 4433-4440. DOI: 10.1128/AAC.00334-09
Yen, H., Hoffmann, E., Taylor, G., Scholtissek, C., Monto, A., Webster, R., & Govorkova, E. (2006) Importance of Neuraminidase Active-Site Residues to the Neuraminidase Inhibitor Resistance of Influenza Viruses. Journal of Virology, 80(17), 8787-8795. DOI: 10.1128/jvi.00477-06
by Clay Clark in Biochem Blogs
Many scientists out there want to know about the dynamics of a protein or how a protein binds to small molecules. But sometimes that information is hard to get using classical techniques such as X-ray crystallography or NMR. Maybe … Continue reading →... Read more »
Zhang, X., Chien, E., Chalmers, M., Pascal, B., Gatchalian, J., Stevens, R., & Griffin, P. (2010) Dynamics of the β -Adrenergic G-Protein Coupled Receptor Revealed by Hydrogen−Deuterium Exchange . Analytical Chemistry, 82(3), 1100-1108. DOI: 10.1021/ac902484p
Landgraf, R., Chalmers, M., & Griffin, P. (2011) Automated Hydrogen/Deuterium Exchange Electron Transfer Dissociation High Resolution Mass Spectrometry Measured at Single-Amide Resolution. Journal of The American Society for Mass Spectrometry, 23(2), 301-309. DOI: 10.1007/s13361-011-0298-2
Wales, T., & Engen, J. (2006) Hydrogen exchange mass spectrometry for the analysis of protein dynamics. Mass Spectrometry Reviews, 25(1), 158-170. DOI: 10.1002/mas.20064
by Clay Clark in Biochem Blogs
When nano first met biology by Xun Lu Lysozyme is an enzyme that helps to protect us from getting bacterial infections because it can degrade and utilize the sugars in the bacterial cell wall. A good source of lysozyme … Continue reading →... Read more »
Choi, Y., Moody, I., Sims, P., Hunt, S., Corso, B., Perez, I., Weiss, G., & Collins, P. (2012) Single-Molecule Lysozyme Dynamics Monitored by an Electronic Circuit. Science, 335(6066), 319-324. DOI: 10.1126/science.1214824
by Clay Clark in Biochem Blogs
A newly developed technique using miRNA profiling to characterize human cancer cells... Read more »
Guo, H., Ingolia, N., Weissman, J., & Bartel, D. (2010) Mammalian microRNAs predominantly act to decrease target mRNA levels. Nature, 466(7308), 835-840. DOI: 10.1038/nature09267
Sato, F., Tsuchiya, S., Meltzer, S., & Shimizu, K. (2011) MicroRNAs and epigenetics. FEBS Journal, 278(10), 1598-1609. DOI: 10.1111/j.1742-4658.2011.08089.x
Liu, J., Zheng, M., Tang, Y., Liang, X., & Yang, Q. (2011) microRNAs, an active and versatile group in cancers. International Journal of Oral Science, 3(4), 165-175. DOI: 10.4248/IJOS11063
Lu, J., Getz, G., Miska, E., Alvarez-Saavedra, E., Lamb, J., Peck, D., Sweet-Cordero, A., Ebert, B., Mak, R., Ferrando, A.... (2005) MicroRNA expression profiles classify human cancers. Nature, 435(7043), 834-838. DOI: 10.1038/nature03702
Ramaswamy, S. (2001) Multiclass cancer diagnosis using tumor gene expression signatures. Proceedings of the National Academy of Sciences, 98(26), 15149-15154. DOI: 10.1073/pnas.211566398
by Clay Clark in Biochem Blogs
Traditional light microscopes are not able to resolve images small enough to explore the details of cells. One of the techniques used to investigate nanoscale samples is atomic force microscopy (AFM). AFM uses a very fine tip (atoms in … Continue reading →... Read more »
Fantner, G., Barbero, R., Gray, D., & Belcher, A. (2010) Kinetics of antimicrobial peptide activity measured on individual bacterial cells using high-speed atomic force microscopy. Nature Nanotechnology, 5(4), 280-285. DOI: 10.1038/nnano.2010.29
Carvalho, F., Carneiro, F., Martins, I., Assuncao-Miranda, I., Faustino, A., Pereira, R., Bozza, P., Castanho, M., Mohana-Borges, R., Da Poian, A.... (2011) Dengue Virus Capsid Protein Binding to Hepatic Lipid Droplets (LD) Is Potassium Ion Dependent and Is Mediated by LD Surface Proteins. Journal of Virology, 86(4), 2096-2108. DOI: 10.1128/JVI.06796-11
by Clay Clark in Biochem Blogs
What is referred to as “MudPit” here is not “a pit of mud” but a technique in the mass spectrometry field which stands for “multi-dimensional protein identification technology”, a very powerful approach that has been widely used since the … Continue reading →... Read more »
Washburn, M., Wolters, D., & Yates, J. (2001) Large-scale analysis of the yeast proteome via multidimensional protein identification technology. Nature Biotechnology, 19(3), 242-247. DOI: 10.1038/85686
Yang, F., Shen, Y., Camp, D., & Smith, R. (2012) High-pH reversed-phase chromatography with fraction concatenation for 2D proteomic analysis. Expert Review of Proteomics, 9(2), 129-134. DOI: 10.1586/epr.12.15
Tran, J., Zamdborg, L., Ahlf, D., Lee, J., Catherman, A., Durbin, K., Tipton, J., Vellaichamy, A., Kellie, J., Li, M.... (2011) Mapping intact protein isoforms in discovery mode using top-down proteomics. Nature, 480(7376), 254-258. DOI: 10.1038/nature10575
by Clay Clark in Biochem Blogs
There are a number of processes that lead to the death of a cell: apoptosis, necrosis, and autophagy are the primary mechanisms – each has a distinct biochemical and morphological fingerprint. In a new paper by Brent Stockwell’s lab at … Continue reading →... Read more »
Dixon Scott J., Lemberg Kathryn M., Lamprecht Michael R., Skouta Rachid, Zaitsev Eleina M., Gleason Caroline E., Patel Darpan N., Bauer Andras J., Cantley Alexandra M., & Yang Wan Seok. (2012) Ferroptosis: An Iron-Dependent Form of Nonapoptotic Cell Death. Cell, 149(5), 1072. DOI: 10.1016/j.cell.2012.03.042
Dolma Sonam, Lessnick Stephen L, Hahn William C, & Stockwell Brent R. (2003) Identification of genotype-selective antitumor agents using synthetic lethal chemical screening in engineered human tumor cells. Cancer Cell, 3(3), 296. DOI: 10.1016/S1535-6108(03)00050-3
Yang Wan Seok, & Stockwell Brent R. (2008) Synthetic Lethal Screening Identifies Compounds Activating Iron-Dependent, Nonapoptotic Cell Death in Oncogenic-RAS-Harboring Cancer Cells. Chemistry , 15(3), 245. DOI: 10.1016/j.chembiol.2008.02.010
by Clay Clark in Biochem Blogs
If you’ve taken a biochemistry class, you’ve probably heard the structure-function paradigm for proteins: amino acid sequence dictates how the protein will be folded, and the ordered 3D structure of the protein is necessary for function.(1) For example, proper formation … Continue reading →... Read more »
Dunker A.Keith, Lawson J.David, Brown Celeste J, Williams Ryan M, Romero Pedro, Oh Jeong S, Oldfield Christopher J, Campen Andrew M, Ratliff Catherine M, & Hipps Kerry W. (2001) Intrinsically disordered protein. Journal of Molecular Graphics and Modelling, 19(1), 59. DOI: 10.1016/S1093-3263(00)00138-8
Rautureau Gilles J. P., Day Catherine L., & Hinds Mark G. (2010) Intrinsically Disordered Proteins in Bcl-2 Regulated Apoptosis. International Journal of Molecular Sciences, 11(4), 1824. DOI: 10.3390/ijms11041808
Rodi Diane J, Janes Robert W, Sanganee Hitesh J, Holton Robert A, Wallace B.A, & Makowski Lee. (1999) Screening of a library of phage-displayed peptides identifies human Bcl-2 as a taxol-binding protein. Journal of Molecular Biology, 285(1), 203. DOI: 10.1006/jmbi.1998.2303
by Clay Clark in Biochem Blogs
Normally proteins have a globular shape in order to be enzymatically or structurally relevant. Intrinsically disordered proteins (IDPs) broke the protein norms by maintaining their functional roles with little to no overall structure. Most proteins have regions of disorder, such … Continue reading →... Read more »
Kalkhoven Eric. (2004) CBP and p300: HATs for different occasions. Biochemical Pharmacology, 68(6), 1145-1155. DOI: 10.1016/j.bcp.2004.03.045
Vo N, & Goodman R H. (2001) CREB-binding protein and p300 in transcriptional regulation. The Journal of biological chemistry. PMID: 11279224
Avantaggiati M L, Ogryzko V, Gardner K, Giordano A, Levine A S, & Kelly K. (1997) Recruitment of p300/CBP in p53-dependent signal pathways. Cell, 1175-1184. PMID: 9215639
Ferreon Josephine C, Lee Chul Won, Arai Munehito, Martinez-Yamout Maria A, Dyson H Jane, & Wright Peter E. (2009) Cooperative regulation of p53 by modulation of ternary complex formation with CBP/p300 and HDM2. Proceedings of the National Academy of Sciences of the United States of America. PMID: 19357310
by Clay Clark in Biochem Blogs
When you think of Italy you may conjure up images of fine wines, food hearty enough to suppress the most insatiable appetite or natural beauty only a poet could describe, but I think of a birthplace. The birthplace of an … Continue reading →... Read more »
Alon Uri. (2009) How To Choose a Good Scientific Problem. Molecular Cell, 35(6), 726-728. DOI: 10.1016/j.molcel.2009.09.013
Bourne Philip E., & Friedberg Iddo. (2006) Ten Simple Rules for Selecting a Postdoctoral Position. PLoS Computational Biology, 2(11). DOI: 10.1371/journal.pcbi.0020121
Walters Jad, Schipper Joshua L., Swartz Paul, Mattos Carla, & Clark A. Clay. (2012) Allosteric modulation of caspase 3 through mutagenesis. Bioscience Reports, 32(4), 401-411. DOI: 10.1042/BSR20120037
by Clay Clark in Biochem Blogs
After spending a great deal of time and energy on cancer research everyday, sometimes I like to go home and down a glass of red wine (or two, depending on the day). Then, I am left to ponder the simpler … Continue reading →... Read more »
GAWEL RICHARD. (1998) Red wine astringency: a review. Australian Journal of Grape and Wine Research, 4(2), 74-95. DOI: 10.1111/j.1755-0238.1998.tb00137.x
Pascal Christine, Poncet-Legrand Céline, Imberty Anne, Gautier Catherine, Sarni-Manchado Pascale, Cheynier Véronique, & Vernhet Aude. (2007) Interactions between a Non Glycosylated Human Proline-Rich Protein and Flavan-3-ols Are Affected by Protein Concentration and Polyphenol/Protein Ratio. Journal of Agricultural and Food Chemistry, 55(12), 4895-4901. DOI: 10.1021/jf0704108
Canon Francis, Ballivian Renaud, Chirot Fabien, Antoine Rodolphe, Sarni-Manchado Pascale, Lemoine Jérôme, & Dugourd Philippe. (2011) Folding of a Salivary Intrinsically Disordered Protein upon Binding to Tannins. Journal of the American Chemical Society, 133(20), 7847-7852. DOI: 10.1021/ja200534f
by Clay Clark in Biochem Blogs
Heat shock is the effect of subjecting a cell to a higher temperature than that of the ideal metabolic temperature of the organism. One of the most typical responses of the cells is transcriptional up-regulation of genes encoding heat shock … Continue reading →... Read more »
Paul S., Singh C., Mishra S., & Chaudhuri T. K. (2007) The 69 kDa Escherichia coli maltodextrin glucosidase does not get encapsulated underneath GroES and folds through trans mechanism during GroEL/GroES-assisted folding. The FASEB Journal, 21(11), 2874-2885. DOI: 10.1096/fj.06-7958com
TRUSCOTT Kaye N., HOJ Peter B., & SCOPES Robert K. (1994) Purification and characterization of chaperonin 60 and chaperonin 10 from the anaerobic thermophile Thermoanaerobacter brockii. European Journal of Biochemistry, 222(2), 277-284. DOI: 10.1111/j.1432-1033.1994.tb18866.x
Yamauchi Seiji, Ueda Yuya, Matsumoto Mika, Inoue Umihiko, & Hayashi Hidenori. (2012) Distinct features of protein folding by the GroEL system from a psychrophilic bacterium, Colwellia psychrerythraea 34H. Extremophiles, 16(6), 871-882. DOI: 10.1007/s00792-012-0483-7
by Clay Clark in Biochem Blogs
The cyclotide family is the the largest class of circular proteins with as many as 50,000 predicted members. They are currently only found in the Violaceae, Cucurbitaceae, Rubiaceae and recently (1) Fabaceae family of the plant kingdom (violets, gourds, coffee and legumes, … Continue reading →... Read more »
Poth A. G., Colgrave M. L., Lyons R. E., Daly N. L., & Craik D. J. (2011) From the Cover: Discovery of an unusual biosynthetic origin for circular proteins in legumes. Proceedings of the National Academy of Sciences, 108(25), 10127-10132. DOI: 10.1073/pnas.1103660108
Saska I., Gillon A. D., Hatsugai N., Dietzgen R. G., Hara-Nishimura I., Anderson M. A., & Craik D. J. (2007) An Asparaginyl Endopeptidase Mediates in Vivo Protein Backbone Cyclization. Journal of Biological Chemistry, 282(40), 29721-29728. DOI: 10.1074/jbc.M705185200
by Clay Clark in Biochem Blogs
In our Protein Journal Club this semester, we are studying proteins from extremophiles. As their name suggests, extremophiles are organisms that can survive under extreme conditions. These extreme conditions include acidic or basic environments, severe hot or cold environments, lack of … Continue reading →... Read more »
Aghajari Nushin, Haser Richard, Feller Georges, & Gerday Charles. (1998) Crystal structures of the psychrophilic α-amylase from Alteromonas haloplanctis in its native form and complexed with an inhibitor. Protein Science, 7(6), 1481-1481. DOI: 10.1002/pro.5560070626
Aghajari Nushin, Feller Georges, Gerday Charles, & Haser Richard. (1998) Structures of the psychrophilic Alteromonas haloplanctis α-amylase give insights into cold adaptation at a molecular level. Structure, 6(12), 1503-1516. DOI: 10.1016/S0969-2126(98)00149-X
Feller Georges. (2010) Protein stability and enzyme activity at extreme biological temperatures. Journal of Physics: Condensed Matter, 22(32), 323101. DOI: 10.1088/0953-8984/22/32/323101
Gilis Dimitri. (2006) In Silico Analysis of the Thermodynamic Stability Changes of Psychrophilic and Mesophilic α-Amylases upon Exhaustive Single-Site Mutations. ChemInform, 37(31). DOI: 10.1002/chin.200631215
by Clay Clark in Biochem Blogs
The Gibbs Conference on Biothermodynamics recently held its 26th annual meeting in Carbondale, IL. The Gibbs conference began in 1986 with a meeting to discuss the discipline of thermodynamics in biological systems. How does one apply the rigorous techniques utilized … Continue reading →... Read more »
Ackers Gary K., & Bolen D.Wayne. (1997) The Gibbs conference on biothermodynamics: Origins and evolution. Biophysical Chemistry, 64(1-3), 5. DOI: 10.1016/S0301-4622(96)02246-6
Vogt Austin D., & Di Cera Enrico. (2012) Conformational Selection or Induced Fit? A Critical Appraisal of the Kinetic Mechanism. Biochemistry, 51(30), 5902. DOI: 10.1021/bi3006913
Shea Madeline A., Correia John J., & Brenowitz Michael D. (2011) Introduction: Twenty five years of the Gibbs Conference on Biothermodynamics. Biophysical Chemistry, 159(1), 5. DOI: 10.1016/j.bpc.2011.07.002
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