Biochemistry Commons^™

Laser-Induced Photon-Branched Chain Reaction In A Chemically-Active Gas-Dispersed Medium, Thomas George, Renat Letfullin, Galen Duree

Chemistry & Biochemistry Faculty Works

A promising avenue in the development of high-energy pulsed chemical HF/DF lasers and amplifiers is the utilization of a photon-branched chain reaction initiated in a two-phase active medium, that is, a medium containing a laser working gas and ultradispersed passivated metal particles. These particles are evaporated under the action of IR laser radiation which results in the appearance of free atoms, their diffusion into the gas, and the development of a photon-branching chain process, which involves photons as both reactants and products. The key obstacle here is the formation of a relatively large volume (in excess of 10^3 cm^3) of …

Structure And Dynamics Of Metalloproteins In Live Cells, Jeremy D. Cook, James E. Penner-Hahn, Timothy L. Stemmler

Biochemistry and Molecular Biology Faculty Publications

X-ray absorption spectroscopy (XAS) has emerged as one of the premier tools for investigating the structure and dynamic properties of metals in cells and in metal containing biomolecules. Utilizing the high flux and broad energy range of X-rays supplied by synchrotron light sources, one can selectively excite core electronic transitions in each metal. Spectroscopic signals from these electronic transitions can be used to dissect the chemical architecture of metals in cells, in cellular components and in biomolecules at varying degrees of structural resolution. With the development of ever-brighter X-ray sources, X-ray methods have grown into applications that can be utilized …

The Effect Of Oxidant And The Non-Oxidant Alteration Of Cellular Thiol Concentration On The Formation Of Protein Mixed-Disulfides In Hek 293 Cells, Jasen Lee Gilge, Michael Fisher, Yuh-Cherng Chai

Chemistry

Cellular molecules possess various mechanisms in responding to oxidant stress. In terms of protein responses, protein S-glutathionylation is a unique post-translational modification of protein reactive cysteines forming disulfides with glutathione molecules. This modification has been proposed to play roles in antioxidant, regulatory and signaling in cells under oxidant stress. Recently, the increased level of protein S-glutathionylation has been linked with the development of diseases. In this report, specific S-glutathionylated proteins were demonstrated in human embryonic kidney 293 cells treated with two different oxidative reagents: diamide and hydrogen peroxide. Diamide is a chemical oxidizing agent whereas hydrogen peroxide is a physiological …

Three-Dimensional Microfluidic Devices Fabricated In Layered Paper And Tape, Andres W. Martinez, Scott T. Phillips, George M. Whitesides

Chemistry and Biochemistry

This article describes a method for fabricating 3D microfluidic devices by stacking layers of patterned paper and double-sided adhesive tape. Paper-based 3D microfluidic devices have capabilities in microfluidics that are difficult to achieve using conventional open-channel microsystems made from glass or polymers. In particular, 3D paper-based devices wick fluids and distribute microliter volumes of samples from single inlet points into arrays of detection zones (with numbers up to thousands). This capability makes it possible to carry out a range of new analytical protocols simply and inexpensively (all on a piece of paper) without external pumps. We demonstrate a prototype 3D …

Sinorhizobium Meliloti, A Bacterium Lacking The Autoinducer-2 (Ai-2) Synthase, Responds To Ai-2 Supplied By Other Bacteria, C. S. Pereira, J. R. Mcauley, M. E. Taga, K. B. Xavier, Stephen T. Miller

Chemistry & Biochemistry Faculty Works

Many bacterial species respond to the quorum-sensing signal autoinducer-2 (AI-2) by regulating different niche-specific genes. Here, we show that Sinorhizobium meliloti, a plant symbiont lacking the gene for the AI-2 synthase, while not capable of producing AI-2 can nonetheless respond to AI-2 produced by other species. We demonstrate that S. meliloti has a periplasmic binding protein that binds AI-2. The crystal structure of this protein (here named SmlsrB) with its ligand reveals that it binds (2R,4S)-2-methyl-2,3,3,4-tetrahydroxytetrahydrofuran (R-THMF), the identical AI-2 isomer recognized by LsrB of Salmonella typhimurium. The gene encoding SmlsrB is in an operon with orthologues of the lsr …

Characterization Of The Ompl1 Gene Of Pathogenic Leptospira Species In China And Cross-Immunogenicity Of The Ompl1 Protein, Haiyan Dong, Ye Hu, Feng Xue, Dexter Sun, David M. Ojcius, Yafei Mao, Jie Yan

All Dugoni School of Dentistry Faculty Articles

Background: The usefulness of available vaccine and serological tests for leptospirosis is limited by the low cross-reactivity of antigens from numerous serovars of pathogenic Leptospira spp. Identification of genus-specific protein antigens (GP-Ag) of Leptospira would be important for development of universal vaccines and serodiagnostic methods. OmpL1, a transmembrane porin of pathogenic leptospires, was identified as a possible GP-Ag, but its sequence diversity and immune cross-reactivity among different serovars of pathogenic leptospires remains largely unknown.

Results: PCR analysis demonstrated that the ompL1 gene existed in all 15 official Chinese standard strains as well as 163 clinical strains of pathogenic leptospires isolated …

Evidence Of A Novel Rna Secondary Structure In The Coding Region Of Hiv-1 Pol Gene., Qi Wang, Ian Barr, Feng Guo, Christopher Lee

Natural Sciences and Mathematics | Faculty Scholarship

RNA secondary structures play several important roles in the human immunodeficiency virus (HIV) life cycle. To assess whether RNA secondary structure might affect the function of the HIV protease and reverse transcriptase genes, which are the main targets of anti-HIV drugs, we applied a series of different computational approaches to detect RNA secondary structures, including thermodynamic RNA folding predictions, synonymous variability analysis, and covariance analysis. Each method independently revealed strong evidence of a novel RNA secondary structure at the junction of the protease and reverse transcriptase genes, consisting of a 107-nucleotide region containing three stems, A, B, and C. First, …

Comparison Of Electrically Mediated And Liposome-Complexed Plasmid Dna Delivery To The Skin, Loree C. Heller, Mark J. Jaroszeski, Domenico Coppola, Richard Heller

Bioelectrics Publications

BACKGROUND: Electroporation is an established technique for enhancing plasmid delivery to many tissues in vivo, including the skin. We have previously demonstrated efficient delivery of plasmid DNA to the skin utilizing a custom-built four-plate electrode. The experiments described here further evaluate cutaneous plasmid delivery using in vivo electroporation. Plasmid expression levels are compared to those after liposome mediated delivery.

METHODS: Enhanced electrically-mediated delivery, and less extensively, liposome complexed delivery, of a plasmid encoding the reporter luciferase was tested in rodent skin. Expression kinetics and tissue damage were explored as well as testing in a second rodent model.

RESULTS: Experiments …

The Proton-Translocating A Subunit Of F0f1-Atp Synthase Is Allocated Asymmetrically To The Peripheral Stalk., Monika G Düser, Yumin Bi, Nawid Zarrabi, Stanley D Dunn, Michael Börsch

Biochemistry Publications

The position of the a subunit of the membrane-integral F0 sector of Escherichia coli ATP synthase was investigated by single molecule fluorescence resonance energy transfer studies utilizing a fusion of enhanced green fluorescent protein to the C terminus of the a subunit and fluorescent labels attached to specific positions of the epsilon or gamma subunits. Three fluorescence resonance energy transfer levels were observed during rotation driven by ATP hydrolysis corresponding to the three resting positions of the rotor subunits, gamma or epsilon, relative to the a subunit of the stator. Comparison of these positions of the rotor sites with those …

Domain Compliance And Elastic Power Transmission In Rotary F(O)F(1)-Atpase., Hendrik Sielaff, Henning Rennekamp, André Wächter, Hao Xie, Florian Hilbers, Katrin Feldbauer, Stanley D Dunn, Siegfried Engelbrecht, Wolfgang Junge

Biochemistry Publications

The 2 nanomotors of rotary ATP synthase, ionmotive F(O) and chemically active F(1), are mechanically coupled by a central rotor and an eccentric bearing. Both motors rotate, with 3 steps in F(1) and 10-15 in F(O). Simulation by statistical mechanics has revealed that an elastic power transmission is required for a high rate of coupled turnover. Here, we investigate the distribution in the F(O)F(1) structure of compliant and stiff domains. The compliance of certain domains was restricted by engineered disulfide bridges between rotor and stator, and the torsional stiffness (kappa) of unrestricted domains was determined by analyzing their thermal rotary …

Cariporide Prodrugs: Targeting Brain Cancer Cells Through Sodium-Proton Exchange Inhibition, Jacob Vervynckt, Johnathan Brantley, Jessica Moore, Mark Graves Ii, William Harley, Fredric A. Gorin, Hasan Palandoken

Chemistry and Biochemistry

More than 200,000 people in the United States are diagnosed with a primary or metastatic brain tumor annually. The life expectancy for these individuals is approximately 9-12 months from the time of diagnosis. This poor prognosis is due to the ineffectiveness of existing therapies (i.e., chemotherapy and radiotherapy) against brain cancer, where the primary problem is the inability to differentiate cancer cells from healthy brain cells.

Relative to healthy brain tissue, the heightened metabolism of cancer cells increases their reliance on the ion transport proteins NHE (sodium-proton exchanger) and NCX (sodium-calcium exchanger). Inhibition of these proteins disrupts the intricate pH …

Alkoxyamine Polymers: Versatile Materials For Surface Ligation Applications, Lindsey Hines, Jessica Moore, David Dahl, Belinda Lady, Johnathan Brantley, Hasan Palandoken

Chemistry and Biochemistry

Immobilization of biomolecules (i.e., proteins, carbohydrates), on polymeric surfaces has been an area of intense research. The resultant bioconjugates often display increased stability, bioavailability and activity. Our research program seeks to explore the utility of the alkoxyamine (RONH2) functional group in new materials as versatile ligating sites for the immobilization of various compounds.

The ease with which alkoxyamines (RONH2) condense with aldehydes or ketones has prompted their widespread use in labelling liposome, bacterial and mammalian cell surfaces as well as chemoselectively ligating small molecule ‘recognition elements' onto polyfunctional substrates. These condensation reactions proceed in aqueous media to afford the robust …

Removal Of Benzaldehyde From Methanol Using Alkoxyamine Functionalized Silica Gel, Dino Sulejmanovic, Carrie Jo Pruitt, Eric Conte, Hasan Palandoken, Shing-Yi Suen

Chemistry and Biochemistry

Alkoxyamines (RONH₂) react readily with aldehydes and ketones to form stable oxime ethers, and, thus are expected to be excellent scavengers of these compounds. We installed alkoxyamines on silica gel surface to remove benzaldehyde from methanol. Silica gel was immersed in H₂O₂/H₂SO₄ solution to activate the surface. Activated silica gel was then reacted with 3-chloropropyltrichlorosilane. Displacement of the chloride with N-hydroxyphthalimide and subsequent treatment with hydrazine provided alkoxyamines on the surface of silica gel. The resulting silica gel surface with alkoxyamine functional groups was immersed in a 100 ppm benzaldehyde solution …

Molecular Characterisation Of A Bovine-Like Rotavirus Detected From A Giraffe, Emily Mulherin, Jill Bryan, Marijke Beltman, Luke O'Grady, Eugene Pidgeon, Lucie Garon, Andrew Lloyd, John Bainbridge, Helen O'Shea, Paul Whyte, Séamus Fanning

Department of Biological Sciences Publications

Background

Rotavirus (RV), is a member of the Reoviridae family and an important etiological agent of acute viral gastroenteritis in the young. Rotaviruses have a wide host range infecting a broad range of animal species, however little is known about rotavirus infection in exotic animals. In this paper we report the first characterisation of a RV strain from a giraffe calf.

Results

This report describes the identification and detailed molecular characterisation of a rotavirus strain detected from a 14-day-old Giraffe (Giraffa camelopardalis), presenting with acute diarrhea. The RV strain detected from the giraffe was characterized molecularly as G10P[11]. …

The Effects Of A Networked Data Acquisition System On Student Learning In General Chemistry Laboratories, Stuart C. Burris, Les L. Pesterfield, Darwin Dahl, Hasan Palandoken, Jacqueline Pope-Tarrence

Chemistry and Biochemistry

We have recently incorporated a networked data acquisition and analysis system into in our second semester general chemistry laboratories. Our investigation team has conducted a study to evaluate the effects of the networked data acquisition system on student performance on a Beer's Law laboratory. We focused on two specific phenomena: the effect of the networked data acquisition system on 1) student learning of specific content material and 2) student comfort level with laboratory work and data analysis. The study involved a networked data acquisition group (employing a UV-Vis spectrometer and colorimeters) and a control group (employing traditional, single wavelength spectrometers). …

Tethering Polypeptides Through Bifunctional Peg Cross-Linking Agents To Probe Protein Function: Application To Atp Synthase., Daniel J Cipriano, Stanley D Dunn

Biochemistry Publications

Chemical crosslinking mediated by short bifunctional reagents has been widely used for determining physical relationships among polypeptides in multisubunit proteins, but less often for functional studies. Here we introduce the approach of tethering polypeptides by using bifunctional reagents containing a lengthy, flexible PEG linker as a form of crosslinking especially suited to functional analyses. The rotary molecular motor ATP synthase was used as a model subject. Single cysteine residues were introduced into selected positions of ATP synthase epsilon subunit, a component of the rotor subcomplex of the enzyme, and the unrelated maltose binding protein (MBP), then the two purified recombinant …

An Inositolphosphorylceramide Synthase Is Involved In Regulation Of Plant Programmed Cell Death Associated With Defense In Arabidopsis, Wenming Wang, Xiaohua Yang, Samantha Tangchiaburana, Roland Ndeh, Jennifer E. Markham, Yoseph Tsegaye, Teresa M. Dunn, Guo-Liang Wang, Maria Bellizzi, James F. Parsons, Danielle Morrissey, Janis E. Bravo, Daniel V. Lynch, Shunyuan Xiao

Department of Biochemistry: Faculty Publications

The Arabidopsis thaliana resistance gene RPW8 triggers the hypersensitive response (HR) to restrict powdery mildew infection via the salicylic acid–dependent signaling pathway. To further understand how RPW8 signaling is regulated, we have conducted a genetic screen to identify mutations enhancing RPW8-mediated HR-like cell death (designated erh). Here, we report the isolation and characterization of the Arabidopsis erh1 mutant, in which the At2g37940 locus is knocked out by a T-DNA insertion. Loss of function of ERH1 results in salicylic acid accumulation, enhanced transcription of RPW8 and RPW8-dependent spontaneous HR-like cell death in leaf tissues, and reduction in plant stature. …

Global Analysis Of Gene Expression Changes During Retinoic Acid-Induced Growth Arrest And Differentiation Of Melanoma: Comparison To Differentially Expressed Genes In Melanocytes Vs Melanoma, Mary H. Estler, Goran Boskovic, James Denvir, Sarah Miles, Donald A. Primerano, Richard M. Niles

Biochemistry and Microbiology

BACKGROUND: The incidence of malignant melanoma has significantly increased over the last decade. Some of these malignancies are susceptible to the growth inhibitory and pro-differentiating effects of all-trans-retinoic acid (RA). The molecular changes responsible for the biological activity of RA in melanoma are not well understood.

RESULTS: In an analysis of sequential global gene expression changes during a 4-48 h RA treatment of B16 mouse melanoma cells, we found that RA increased the expression of 757 genes and decreased the expression of 737 genes. We also compared the gene expression profile (no RA treatment) between non-malignant melan-a mouse melanocytes and …

Simple Inhibitors Of Histone Deacetylase Activity That Combine Features Of Short-Chain Fatty Acid And Hydroxamic Acid Inhibitors, Jessica L. Tischler, Basel Abuaita, Sierra C. Cuthpert, Christopher Fage, Kristi Murphy, Andrew Saxe, Edward B. Furr, Jamie Hedrick, Jennifer Meyers, David Snare, Ali R. Zand

Chemistry & Biochemistry Publications

Butyric acid and trichostatin A (TSA) are anti-cancer compounds that cause the upregulation of genes involved in differentiation and cell cycle regulation by inhibiting histone deacetylase (HDAC) activity. In this study we have synthesized and evaluated compounds that combine the bioavailability of short-chain fatty acids, like butyric acid, with the bidentate binding ability of TSA. A series of analogs were made to examine the effects of chain length, simple aromatic cap groups, and substituted hydroxamates on the compounds' ability to inhibit rat-liver HDAC using a fluorometric assay. In keeping with previous structure-activity relationships, the most effective inhibitors consisted of longer …

Molecular-Orientation-Dependent Ac Stark Effect And Its Impact On Multiphoton Processes, Xi Chu

Chemistry and Biochemistry Faculty Publications

We study the dependence of the ac Stark shifts of electronic energies on the molecular orientation relative to the polarization direction of an incident intense laser field, using a three-dimensional non-Hermitian Floquet method and H(2)+ as a model system. Simultaneously, we also study the orientation-dependent high-order harmonic generation (HHG) and multiphoton ionization (MPI). We find that with the presence of near-one-photon resonance, the Stark effect strongly mixes electronic states of different symmetries to create quasienergy states (QESs). The orientation dependence of multiphoton processes, in which these QESs play an important role, becomes complex. Population transfer is better achieved with aligned …

Phosphorylation Of P27kip1 Regulates Assembly And Activation Of Cyclin D1-Cdk4, Michelle D. Larrea, Jiyong Liang, Thiago G. Da Silva, Feng Hong, Shan H. Shao, Kathy Han, D. Dumont, Joyce M. Slingerland

Chemistry and Physics Faculty Articles

p27 mediates Cdk2 inhibition and is also found in cyclin D1-Cdk4 complexes. The present data support a role for p27 in the assembly of D-type cyclin-Cdk complexes and indicate that both cyclin D1-Cdk4-p27 assembly and kinase activation are regulated by p27 phosphorylation. Prior work showed that p27 can be phosphorylated by protein kinase B/Akt (PKB/Akt) at T157 and T198. Here we show that PKB activation and the appearance of p27pT157 and p27pT198 precede p27-cyclin D1-Cdk4 assembly in early G₁. PI3K/PKB inhibition rapidly reduced p27pT157 and p27pT198 and dissociated cellular p27-cyclin D1-Cdk4. Mutant p27 allele products lacking phosphorylation at …

Mutational Optimization Of The Coelenterazine-Dependent Luciferase From Renilla, Jongchan Woo, Albrecht G. Von Arnim

Faculty Publications and Other Works -- Biochemistry, Cellular and Molecular Biology

Renilla luciferase (RLUC) is a popular reporter enzyme for gene expression and biosensor applications, but it is an unstable enzyme whose catalytic mechanism remains to be elucidated. We titrated that one RLUC molecule can turn over about one hundred molecules of coelenterazine substrate. Mutagenesis of active site residue Pro220 extended the half-life of photon emission, yielding brighter luminescence in E. coli. Random mutagenesis uncovered two new mutations that stabilized and increased photon emission in vivo and in vitro, while ameliorating substrate inhibition. Further amended with a previously identified mutation, a new triple mutant showed a threefold improved k …

The Very-Long-Chain Hydroxy Fatty Acyl-Coa Dehydratase Pasticcino2 Is Essential And Limiting For Plant Development, Liên Bach, Louise V. Michaelson, Richard Haslam, Yannick Bellec, Lionel Gissot, Jessica Marion, Marco Da Costa, Jean-Pierre Boutin, Martine Miquel, Frédérique Tellier, Frederic Domergue, Jennifer E. Markham, Frederic Beaudoin, Johnathan A. Napier, Jean-Denis Faure

Department of Biochemistry: Faculty Publications

Very-long-chain fatty acids (VLCFAs) are synthesized as acyl-CoAs by the endoplasmic reticulum-localized elongase multiprotein complex. Two Arabidopsis genes are putative homologues of the recently identified yeast 3-hydroxy-acyl-CoA dehydratase (PHS1), the third enzyme of the elongase complex. We showed that Arabidopsis PASTICCINO2 (PAS2) was able to restore phs1 cytokinesis defects and sphingolipid long chain base overaccumulation. Conversely, the expression of PHS1 was able to complement the developmental defects and the accumulation of long chain bases of the pas2–1 mutant. The pas2–1 mutant was characterized by a general reduction of VLCFA pools in seed storage triacylglycerols, cuticular waxes, …

Oriented Cell Growth On Self-Assembled Bacteriophage M13 Thin Films, Jianhua Rong, L. Andrew Lee, Kai Li, Brandon Harp, Charlene M. Mello, Zhongwei Niu, Qian Wang

Faculty Publications

Fibrillar M13 bacteriophages were used as basic building blocks to generate thin films with aligned nanogrooves, which, upon chemical grafting with RGD peptides, guide cell alignment and orient the cell outgrowth along defined directions.

Sex- And Gonad-Affecting Scent Compounds And 3 Male Pheromones In The Rat, Jian-Xu Zhang, Lixing Sun, Jin-Hua Zhang, Zhi-Yong Feng

All Faculty Scholarship for the College of the Sciences

This study was aimed at identifying sex pheromones of the rat (Rattus norvegicus). We characterized the volatiles and semivolatiles of rat preputial gland and voided urine by using gas chromatography–mass spectrometry (GC–MS) and quantified them by their GC areas (abundances) and percentage of GC areas (relative abundances). Although all the compounds other than 4-heptanone and phenol detected were shared by males and females, the quantities for some of these sex-common compounds exhibited sexual dimorphism and decreased with gonadectomy. Thus, these compounds might be sex pheromones. Among them, squalene from preputial glands and 2-heptanone and 4-ethyl phenol from urine …

Spectroscopic Studies On Arabidopsis Ethe1, A Glyoxalase Ii-Like Protein, Meghan M. Holdorf, Brian Bennett, Michael W. Crowder, Christopher A. Makaroff

Physics Faculty Research and Publications

ETHE1 (ethylmalonic encephalopathy protein 1) is a β-lactamase fold-containing protein that is essential for the survival of a range of organisms. In spite of the apparent importance of this enzyme, very little is known about its function or biochemical properties. In this study Arabidopsis ETHE1 was over-expressed and purified and shown to bind tightly to 1.2 ± 0.2 equivalents of iron. ¹H NMR and EPR studies demonstrate that the predominant oxidation state of Fe in ETHE1 is Fe(II), and NMR studies confirm that two histidines are bound to Fe(II). EPR studies show that there is no antiferromagnetically coupled Fe(III)Fe(II) …

Small Heat Shock Protein Activity Is Regulated By Variable Oligomeric Substructure, J. L. Benesch, M. Ayoub, C. V. Robinson, J. A. Aquilina

Faculty of Science - Papers (Archive)

The alpha-crystallins are members of the small heat shock protein (sHSP) family of molecular chaperones which have evolved to minimize intracellular protein aggregation, however they are also implicated in a number of protein deposition diseases. In this study we have employed novel mass spectrometry techniques to investigate the changes in quaternary structure associated with this switch from chaperone to adjuvant of aggregation. We have replicated the oligomeric rearrangements observed for in vivo disease-related modifications, without altering the protein sequence, by refolding the alpha-crystallins in vitro. This refolding results in a loss of dimeric substructure concomitant with an augmentation of substrate …

Altered Expression Of Cell Cycle And Apoptotic Proteins In Chronic Hepatitis C Virus Infection., Saira Sarfraz, Saeed Hamid, Anwar Ali Siddiqui, Snawar Hussain, Shahid Pervez, Graeme Alexander

Department of Biological & Biomedical Sciences

Background: A disrupted cell cycle progression of hepatocytes was reported in chronic hepatitis C virus (HCV) infection, which can contribute significantly in the associated pathogenesis. The present study aimed to further elaborate these disruptions by evaluating the expression of key cell cycle and apoptotic proteins in chronic HCV infection with particular reference to genotype 3. Archival liver biopsy specimens of chronic HCV-infection (n = 46) and normal histology (n = 5) were analyzed by immunohistochemistry using antibodies against proliferation marker Mcm-2, G1 phase marker Cyclin D1, S phase marker Cyclin A, cell cycle regulators p21 (CDK inhibitor) and p53 (tumor …

Spatial And Temporal Patterns In The Microbial Diversity Of A Meromictic Soda Lake In Washington State, Pedro A. Dimitriu, Holly C. Pinkart, Brent M. Peyton, Melanie R. Mormile

All Faculty Scholarship for the College of the Sciences

The microbial community diversity and composition of meromictic Soap Lake were studied using culture-dependent and culture-independent approaches. The water column and sediments were sampled monthly for a year. Denaturing gradient gel electrophoresis of bacterial and archaeal 16S rRNA genes showed an increase in diversity with depth for both groups. Late-summer samples harbored the highest prokaryotic diversity, and the bacteria exhibited less seasonal variability than the archaea. Most-probable-number assays targeting anaerobic microbial guilds were performed to compare summer and fall samples. In both seasons, the anoxic samples appeared to be dominated by lactate-oxidizing sulfate-reducing prokaryotes. High numbers of lactate- and acetate-oxidizing …

Probing The Functional Tolerance Of The B Subunit Of Escherichia Coli Atp Synthase For Sequence Manipulation Through A Chimera Approach., Yumin Bi, Joel C Watts, Pamela Krauss Bamford, Lee-Ann K Briere, Stanley D Dunn

Biochemistry Publications

A dimer of 156-residue b subunits forms the peripheral stator stalk of eubacterial ATP synthase. Dimerization is mediated by a sequence with an unusual 11-residue (hendecad) repeat pattern, implying a right-handed coiled coil structure. We investigated the potential for producing functional chimeras in the b subunit of Escherichia coli ATP synthase by replacing parts of its sequence with corresponding regions of the b subunits from other eubacteria, sequences from other polypeptides having similar hendecad patterns, and sequences forming left-handed coiled coils. Replacement of positions 55-110 with corresponding sequences from Bacillus subtilis and Thermotoga maritima b subunits resulted in fully functional …