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Full-Text Articles in Medicine and Health Sciences

Synthesis And Processing Of The Alpha Heavy Chains Of Secreted And Membrane-Bound Iga, H. Kikutani, R. Sitia, R. A. Good, Janet Stavnezer Apr 2008

Synthesis And Processing Of The Alpha Heavy Chains Of Secreted And Membrane-Bound Iga, H. Kikutani, R. Sitia, R. A. Good, Janet Stavnezer

Janet M. Stavnezer

We have compared the synthesis and processing of immunoglobulin alpha chains in two murine cell lines, a B cell lymphoma that expresses membrane-bound IgA and a hybridoma that secretes IgA. Results of biosynthetic labeling experiments demonstrated that membrane-bound and secreted alpha chains have two distinct intracellular precursors, of different molecular weights and isoelectric points. RNAs from both of these cell lines direct the synthesis in vitro of two alpha polypeptides of Mr 59,000 and 62,000, the larger one being the precursor for membrane-bound alpha chain and the smaller one being the precursor for secreted alpha chain. These cell lines each …


Dictyostelium Discoideum Plasma Membranes Contain An Actin-Nucleating Activity That Requires Ponticulin, An Integral Membrane Glycoprotein, A. Shariff, Elizabeth J. Luna Mar 2008

Dictyostelium Discoideum Plasma Membranes Contain An Actin-Nucleating Activity That Requires Ponticulin, An Integral Membrane Glycoprotein, A. Shariff, Elizabeth J. Luna

Elizabeth J. Luna

In previous equilibrium binding studies, Dictyostelium discoideum plasma membranes have been shown to bind actin and to recruit actin into filaments at the membrane surface. However, little is known about the kinetic pathway(s) through which actin assembles at these, or other, membranes. We have used actin fluorescently labeled with N-(1-pyrenyl)iodoacetamide to examine the kinetics of actin assembly in the presence of D. discoideum plasma membranes. We find that these membranes increase the rate of actin polymerization. The rate of membrane-mediated actin polymerization is linearly dependent on membrane protein concentrations up to 20 micrograms/ml. Nucleation (the association of activated actin monomers …


Ponticulin Plays A Role In The Positional Stabilization Of Pseudopods, D. C. Shutt, D. Wessels, K. Wagenknecht, A. Chandrasekhar, Anne L. Hitt, Elizabeth J. Luna, D. R. Soll Mar 2008

Ponticulin Plays A Role In The Positional Stabilization Of Pseudopods, D. C. Shutt, D. Wessels, K. Wagenknecht, A. Chandrasekhar, Anne L. Hitt, Elizabeth J. Luna, D. R. Soll

Elizabeth J. Luna

Ponticulin is a 17-kD glycoprotein that represents a major high affinity link between the plasma membrane and the cortical actin network of Dictyostelium. To assess the role of ponticulin in pseudopod extension and retraction, the motile behavior of two independently generated mutants lacking ponticulin was analyzed using computer-assisted two- and three-dimensional motion analysis systems. More than half of the lateral pseudopods formed off the substratum by ponticulin-minus cells slipped relative to the substratum during extension and retraction. In contrast, all pseudopods formed off the substratum by wild-type cells were positionally fixed in relation to the substratum. Ponticulin-minus cells also formed …


A Membrane Cytoskeleton From Dictyostelium Discoideum. Ii. Integral Proteins Mediate The Binding Of Plasma Membranes To F-Actin Affinity Beads, Elizabeth J. Luna, C. M. Goodloe-Holland, H. M. Ingalls Mar 2008

A Membrane Cytoskeleton From Dictyostelium Discoideum. Ii. Integral Proteins Mediate The Binding Of Plasma Membranes To F-Actin Affinity Beads, Elizabeth J. Luna, C. M. Goodloe-Holland, H. M. Ingalls

Elizabeth J. Luna

In novel, low-speed sedimentation assays, highly purified, sonicated Dictyostelium discoideum plasma membrane fragments bind to F-actin beads (fluorescein-labeled F-actin on antifluorescein IgG-Sephacryl S-1000 beads). Binding was found to be (a) specific, since beads containing bound fluorescein-labeled ovalbumin or beads without bound fluorescein-labeled protein do not bind membranes, (b) saturable at approximately 0.6 microgram of membrane protein per microgram of bead-bound F-actin, (c) rapid with a t1/2 of 4-20 min, and (d) apparently of reasonable affinity since the off rate is too slow to be measured by present techniques. Using low-speed sedimentation assays, we found that sonicated plasma membrane fragments, after …


Binding And Assembly Of Actin Filaments By Plasma Membranes From Dictyostelium Discoideum, M. A. Schwartz, Elizabeth J. Luna Mar 2008

Binding And Assembly Of Actin Filaments By Plasma Membranes From Dictyostelium Discoideum, M. A. Schwartz, Elizabeth J. Luna

Elizabeth J. Luna

The binding of native, 125I-Bolton-Hunter-labeled actin to purified Dictyostelium discoideum plasma membranes was measured using a sedimentation assay. Binding was saturable only in the presence of the actin capping protein, gelsolin. In the presence of gelsolin, the amount of actin bound at saturation to three different membrane preparations was 80, 120, and 200 micrograms/mg of membrane protein. The respective concentrations of actin at half-saturation were 8, 12, and 18 micrograms/ml. The binding curves were sigmoidal, indicating positive cooperativity at low actin concentrations. This cooperativity appeared to be due to actin-actin associations during polymerization, since phalloidin converted the curve to a …


A Membrane Cytoskeleton From Dictyostelium Discoideum. I. Identification And Partial Characterization Of An Actin-Binding Activity, Elizabeth J. Luna, V. M. Fowler, J. Swanson, D. Branton, D. L. Taylor Mar 2008

A Membrane Cytoskeleton From Dictyostelium Discoideum. I. Identification And Partial Characterization Of An Actin-Binding Activity, Elizabeth J. Luna, V. M. Fowler, J. Swanson, D. Branton, D. L. Taylor

Elizabeth J. Luna

Dictyostelium discoideum plasma membranes isolated by each of three procedures bind F-actin. The interactions between these membranes and actin are examined by a novel application of falling ball viscometry. Treating the membranes as multivalent actin-binding particles analogous to divalent actin-gelation factors, we observe large increases in viscosity (actin cross-linking) when membranes of depleted actin and myosin are incubated with rabbit skeletal muscle F-actin. Pre-extraction of peripheral membrane proteins with chaotropes or the inclusion of Triton X-100 during the assay does not appreciably diminish this actin cross-linking activity. Lipid vesicles, heat-denatured membranes, proteolyzed membranes, or membranes containing endogenous actin show minimal …


Supervillin (P205): A Novel Membrane-Associated, F-Actin-Binding Protein In The Villin/Gelsolin Superfamily, Kersi N. Pestonjamasp, Robert K. Pope, J. D. Wulfkuhle, Elizabeth J. Luna Mar 2008

Supervillin (P205): A Novel Membrane-Associated, F-Actin-Binding Protein In The Villin/Gelsolin Superfamily, Kersi N. Pestonjamasp, Robert K. Pope, J. D. Wulfkuhle, Elizabeth J. Luna

Elizabeth J. Luna

Actin-binding membrane proteins are involved in both adhesive interactions and motile processes. We report here the purification and initial characterization of p205, a 205-kD protein from bovine neutrophil plasma membranes that binds to the sides of actin filaments in blot overlays. p205 is a tightly bound peripheral membrane protein that cosediments with endogenous actin in sucrose gradients and immunoprecipitates. Amino acid sequences were obtained from SDS-PAGE-purified p205 and used to generate antipeptide antibodies, immunolocalization data, and cDNA sequence information. The intracellular localization of p205 in MDBK cells is a function of cell density and adherence state. In subconfluent cells, p205 …


Smooth Muscle Archvillin: A Novel Regulator Of Signaling And Contractility In Vascular Smooth Muscle, Samudra S. Gangopadhyay, Norio Takizawa, Cynthia Gallant, Amy L. Barber, Hyun-Dong Je, Tara C. Smith, Elizabeth J. Luna, Kathleen G. Morgan Mar 2008

Smooth Muscle Archvillin: A Novel Regulator Of Signaling And Contractility In Vascular Smooth Muscle, Samudra S. Gangopadhyay, Norio Takizawa, Cynthia Gallant, Amy L. Barber, Hyun-Dong Je, Tara C. Smith, Elizabeth J. Luna, Kathleen G. Morgan

Elizabeth J. Luna

The mechanisms by which protein kinase C (PKC) and extracellular-signal-regulated kinases (ERK1/2) govern smooth-muscle contractility remain unclear. Calponin (CaP), an actin-binding protein and PKC substrate, mediates signaling through ERK1/2. We report here that CaP sequences containing the CaP homology (CH) domain bind to the C-terminal 251 amino acids of smooth-muscle archvillin (SmAV), a new splice variant of supervillin, which is a known actin- and myosin-II-binding protein. The CaP-SmAV interaction is demonstrated by reciprocal yeast two-hybrid and blot-overlay assays and by colocalization in COS-7 cells. In differentiated smooth muscle, endogenous SmAV and CaP co-fractionate and co-translocate to the cell cortex after …


Supervillin Modulation Of Focal Adhesions Involving Trip6/Zrp-1, Norio Takizawa, Tara C. Smith, Thomas Nebl, Jessica Lynn Crowley, Stephen J. Palmieri, Lawrence M. Lifshitz, Anka G. Ehrhardt, Laura M. Hoffman, Mary C. Beckerle, Elizabeth J. Luna Mar 2008

Supervillin Modulation Of Focal Adhesions Involving Trip6/Zrp-1, Norio Takizawa, Tara C. Smith, Thomas Nebl, Jessica Lynn Crowley, Stephen J. Palmieri, Lawrence M. Lifshitz, Anka G. Ehrhardt, Laura M. Hoffman, Mary C. Beckerle, Elizabeth J. Luna

Elizabeth J. Luna

Cell-substrate contacts, called focal adhesions (FAs), are dynamic in rapidly moving cells. We show that supervillin (SV)--a peripheral membrane protein that binds myosin II and F-actin in such cells--negatively regulates stress fibers, FAs, and cell-substrate adhesion. The major FA regulatory sequence within SV (SV342-571) binds to the LIM domains of two proteins in the zyxin family, thyroid receptor-interacting protein 6 (TRIP6) and lipoma-preferred partner (LPP), but not to zyxin itself. SV and TRIP6 colocalize within large FAs, where TRIP6 may help recruit SV. RNAi-mediated decreases in either protein increase cell adhesion to fibronectin. TRIP6 partially rescues SV effects on stress …


Domain Analysis Of Supervillin, An F-Actin Bundling Plasma Membrane Protein With Functional Nuclear Localization Signals, J. D. Wulfkuhle, I. E. Donina, N. H. Stark, Robert K. Pope, Kersi N. Pestonjamasp, M. L. Niswonger, Elizabeth J. Luna Mar 2008

Domain Analysis Of Supervillin, An F-Actin Bundling Plasma Membrane Protein With Functional Nuclear Localization Signals, J. D. Wulfkuhle, I. E. Donina, N. H. Stark, Robert K. Pope, Kersi N. Pestonjamasp, M. L. Niswonger, Elizabeth J. Luna

Elizabeth J. Luna

A growing number of actin-associated membrane proteins have been implicated in motile processes, adhesive interactions, and signal transduction to the cell nucleus. We report here that supervillin, an F-actin binding protein originally isolated from bovine neutrophil plasma membranes, contains functional nuclear targeting signals and localizes at or near vinculin-containing focal adhesion plaques in COS7-2 and CV1 cells. Overexpression of full-length supervillin in these cells disrupts the integrity of focal adhesion plaques and results in increased levels of F-actin and vinculin. Localization studies of chimeric proteins containing supervillin sequences fused with the enhanced green fluorescent protein indicate that: (1) the amino …


The Integral Membrane Protein, Ponticulin, Acts As A Monomer In Nucleating Actin Assembly, C. P. Chia, A. Shariff, S. A. Savage, Elizabeth J. Luna Mar 2008

The Integral Membrane Protein, Ponticulin, Acts As A Monomer In Nucleating Actin Assembly, C. P. Chia, A. Shariff, S. A. Savage, Elizabeth J. Luna

Elizabeth J. Luna

Ponticulin, an F-actin binding transmembrane glycoprotein in Dictyostelium plasma membranes, was isolated by detergent extraction from cytoskeletons and purified to homogeneity. Ponticulin is an abundant membrane protein, averaging approximately 10(6) copies/cell, with an estimated surface density of approximately 300 per microns2. Ponticulin solubilized in octylglucoside exhibited hydrodynamic properties consistent with a ponticulin monomer in a spherical or slightly ellipsoidal detergent micelle with a total molecular mass of 56 +/- 6 kD. Purified ponticulin nucleated actin polymerization when reconstituted into Dictyostelium lipid vesicles, but not when a number of commercially available lipids and lipid mixtures were substituted for the endogenous lipid. …


Archvillin, A Muscle-Specific Isoform Of Supervillin, Is An Early Expressed Component Of The Costameric Membrane Skeleton, Sang W. Oh, Robert K. Pope, Kelly P. Smith, Jessica Lynn Crowley, Thomas Nebl, Jeanne B. Lawrence, Elizabeth J. Luna Mar 2008

Archvillin, A Muscle-Specific Isoform Of Supervillin, Is An Early Expressed Component Of The Costameric Membrane Skeleton, Sang W. Oh, Robert K. Pope, Kelly P. Smith, Jessica Lynn Crowley, Thomas Nebl, Jeanne B. Lawrence, Elizabeth J. Luna

Elizabeth J. Luna

The membrane skeleton protein supervillin binds tightly to both F-actin and membranes and can potentiate androgen receptor activity in non-muscle cells. We report that muscle, which constitutes the principal tissue source for supervillin sequences, contains a approximately 250 kDa isoform of supervillin that localizes within nuclei and with dystrophin at costameres, regions of F-actin membrane attachment in skeletal muscle. The gene encoding this protein, 'archvillin' (Latin, archi; Greek, archos; 'principal' or 'chief'), contains an evolutionarily conserved, muscle-specific 5' leader sequence. Archvillin cDNAs also contain four exons that encode approximately 47 kDa of additional muscle-specific protein sequence in the form of …


F-Actin Binds To The Cytoplasmic Surface Of Ponticulin, A 17-Kd Integral Glycoprotein From Dictyostelium Discoideum Plasma Membranes, L. J. Wuestehube, Elizabeth J. Luna Mar 2008

F-Actin Binds To The Cytoplasmic Surface Of Ponticulin, A 17-Kd Integral Glycoprotein From Dictyostelium Discoideum Plasma Membranes, L. J. Wuestehube, Elizabeth J. Luna

Elizabeth J. Luna

F-actin affinity chromatography and immunological techniques are used to identify actin-binding proteins in purified Dictyostelium discoideum plasma membranes. A 17-kD integral glycoprotein (gp17) consistently elutes from F-actin columns as the major actin-binding protein under a variety of experimental conditions. The actin-binding activity of gp17 is identical to that of intact plasma membranes: it resists extraction with 0.1 N NaOH, 1 mM dithiothreitol (DTT); it is sensitive to ionic conditions; it is stable over a wide range of pH; and it is eliminated by proteolysis, denaturation with heat, or treatment with DTT and N-ethylmaleimide. gp17 may be responsible for much of …


Should Adjustment For Covariates Be Used In Prevalence Estimations, Wenjun Li, Edward J. Stanek, Elizabeth R. Bertone-Johnson Jan 2008

Should Adjustment For Covariates Be Used In Prevalence Estimations, Wenjun Li, Edward J. Stanek, Elizabeth R. Bertone-Johnson

Elizabeth R. Bertone-Johnson

BACKGROUND: Adjustment for covariates (also called auxiliary variables in survey sampling literature) is commonly applied in health surveys to reduce the variances of the prevalence estimators. In theory, adjusted prevalence estimators are more accurate when variance components are known. In practice, variance components needed to achieve the adjustment are unknown and their sample estimators are used instead. The uncertainty introduced by estimating variance components may overshadow the reduction in the variance of the prevalence estimators due to adjustment. We present empirical guidelines indicating when adjusted prevalence estimators should be considered, using gender adjusted and unadjusted smoking prevalence as an illustration. …


Cellular Effects Of Antiepileptic Drugs, Robert L. Macdonald, Michael A. Rogawski Dec 2007

Cellular Effects Of Antiepileptic Drugs, Robert L. Macdonald, Michael A. Rogawski

Michael A. Rogawski

Antiepileptic drugs (AEDs) protect against seizures through interactions with a variety of cellular targets, which include various ion channels, a neurotransmitter transporter, a neurotransmitter metabolic enzyme, and a synaptic vesicle protein. AED actions on these targets can be categorized into four broad groups: 1. Modulation of voltage-dependent ion channels (mainly sodium [Na] but also calcium [Ca] channels) 2. Effects on γ-aminobutyric acid (GABA) systems, including alterations in the cellular disposition of GABA and enhancement of synaptic inhibition mediated by GABA-A receptors 3. Inhibition of synaptic excitation mediated by ionotropic glutamate receptors 4. Modulation of neurotransmitter release, particularly of glutamate, through …


Correlation Network Analysis For Data Integration And Biomarker Selection, Aram Adourian, Ezra Jennings, Raji Balasubramanian, Wade M. Hines, Doris Damian, Tom N. Plasterer, Clary B. Clish, Paul Stroobant, Robert N. Mcburney, Elwin R. Verheij, Ivana I. Bobeldijk, J Van Der Greef, J Lindberg, K Kenne, U Andersson, H Hellmold, K Nilsson, H Salter, I Schuppe-Koistinen Dec 2007

Correlation Network Analysis For Data Integration And Biomarker Selection, Aram Adourian, Ezra Jennings, Raji Balasubramanian, Wade M. Hines, Doris Damian, Tom N. Plasterer, Clary B. Clish, Paul Stroobant, Robert N. Mcburney, Elwin R. Verheij, Ivana I. Bobeldijk, J Van Der Greef, J Lindberg, K Kenne, U Andersson, H Hellmold, K Nilsson, H Salter, I Schuppe-Koistinen

Raji Balasubramanian

This work demonstrates the application of correlation networks to a systems-based investigation of drug-induced hepatotoxicity and the identification of specific and relevant biomarkers in this context.


Effect Of Two Milling Techniques And Flours Particle Size On Some, Theodore N'Dede Djeni Dec 2007

Effect Of Two Milling Techniques And Flours Particle Size On Some, Theodore N'Dede Djeni

Theodore N'Dede Djeni

Two techniques of milling (milling with wooden mortar and milling with hammer grinder) for the production of millet flours were compared in term of physical, biochemical and nutritional aspects and digestibility of the produced flours. A batch of millet grains with candle (Pennisetum glaucum L. R.BR) was purchased from a local market for flour production, while the flour digestibility tests were achieved with an enzymatic crude extract get from gastric juice of and snails (Archachatina marginata). The different rough flours obtained from both milling techniques showed significant differences (p<0.05) in their granulometric profiles and their biochemical composition. The mortar …


Common Pathophysiologic Mechanisms In Migraine And Epilepsy, Michael A. Rogawski Dec 2007

Common Pathophysiologic Mechanisms In Migraine And Epilepsy, Michael A. Rogawski

Michael A. Rogawski

Migraine and epilepsy are comorbid episodic disorders that have common pathophysiologic mechanisms. Migraine attacks, like epileptic seizures, may be triggered by excessive neocortical cellular excitability; in migraine, however, the hyperexcitability is believed to transition to cortical spreading depression rather than to the hypersynchronous activity that characterizes seizures. Some forms of epilepsy and migraine are known to be channelopathies. Mutations in the same genes can cause either migraine or epilepsy or, in some cases, both. Given the likely commonalities in the underlying cellular and molecular mechanisms, it is not surprising that some antiepileptic drugs, including valproate, topiramate, and gabapentin, are effective …


Discordance In The Paraoxonase (Pon1)-192qr Polymorphism, Rozaida @ Poh Yuen Ying Dec 2007

Discordance In The Paraoxonase (Pon1)-192qr Polymorphism, Rozaida @ Poh Yuen Ying

Rozaida @ Poh Yuen Ying

Paraoxonase (PON1) has been implicated to have a cardioprotective role, due to its physical attachment with high-density lipoprotein. PON1192QR is a variation of the PON1 gene, the R allele being a risk factor for cardiovascular disease. Kinetic studies resulting in a plot of paraoxon versus diazoxon hydrolysis rates may be used to accurately predict PON1192 geno-type. In this study, paraoxonase and diazoxonase activities in plasma were measured spec-trophotometrically using plasma while PCR-based PON1192 genotyping was performed us-ing polymerase chain reaction followed by restriction digestion. The two-substrate assay-derived genotypes were cross-referred with those determined by PCR-based genotyping. When results did not …


Functional Generalized Linear Models With Applications To Neuroimaging, Philip T. Reiss, R. Todd Ogden Dec 2007

Functional Generalized Linear Models With Applications To Neuroimaging, Philip T. Reiss, R. Todd Ogden

Philip T. Reiss

No abstract provided.