SYPRO™ Red Protein Gel Stain (5000X in DMSO), 500 μL - Citations
SYPRO™ Red Protein Gel Stain (5000X in DMSO), 500 μL - Citations
View additional product information for SYPRO™ Protein Gel Stains - Citations (S12000, S12001, S6650, S6651, S12010, S12000X3, S6654, S6653, S21900)
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Abstract
Design and characterization of a compact dual channel virus counter.
AuthorsStoffel CL,Kathy RF,Rowlen KL
JournalCytometry. Part A : the journal of the International Society for Analytical Cytology
PubMed ID15830378
Adenosine to inosine editing by ADAR2 requires formation of a ternary complex on the GluR-B R/G site.
AuthorsJaikaran DC, Collins CH, MacMillan AM
JournalJ Biol Chem
PubMed ID12163487
'RNA editing by members of the ADAR (adenosine deaminase that acts on RNA) enzyme family involves hydrolytic deamination of adenosine to inosine within the context of a double-stranded pre-mRNA substrate. Editing of the human GluR-B transcript is catalyzed by the enzyme ADAR2 at the Q/R and R/G sites. We have ... More
Defining the SNARE complex binding surface of alpha-SNAP: implications for SNARE complex disassembly.
AuthorsMarz KE, Lauer JM, Hanson PI
JournalJ Biol Chem
PubMed ID12730228
'N-Ethylmaleimide-sensitive factor (NSF) and its adaptor protein alpha-soluble NSF attachment protein (alpha-SNAP) sustain membrane trafficking by disassembling soluble NSF attachment protein receptor (SNARE) complexes that form during membrane fusion. To better understand the role of alpha-SNAP in this process, we used site-directed mutagenesis to identify residues in alpha-SNAP that interact ... More
Involvement of DnaE, the second replicative DNA polymerase from Bacillus subtilis, in DNA mutagenesis.
AuthorsLe Chatelier E, Bécherel OJ, d'Alençon E, Canceill D, Ehrlich SD, Fuchs RP, Jannière L
JournalJ Biol Chem
PubMed ID14593098
'In a large group of organisms including low G + C bacteria and eukaryotic cells, DNA synthesis at the replication fork strictly requires two distinct replicative DNA polymerases. These are designated pol C and DnaE in Bacillus subtilis. We recently proposed that DnaE might be preferentially involved in lagging strand ... More
Dynamics of myo1c (myosin-ibeta ) lipid binding and dissociation.
AuthorsTang N, Lin T, Ostap EM
JournalJ Biol Chem
PubMed ID12221091
'Myosin-I is the single-headed member of the myosin superfamily that associates with lipid membranes. Biochemical experiments have shown that myosin-I membrane binding is the result of electrostatic interactions between the basic tail domain and acidic phospholipids. To better understand the dynamics of myosin-I membrane association, we measured the rates of ... More
Green-light transilluminator for the detection without photodamage of proteins and DNA labeled with different fluorescent dyes.
AuthorsAlba FJ, Bermúdez A, Daban JR
JournalElectrophoresis
PubMed ID11258745
'The excitation spectra of Nile red and SYPRO red, two currently used dyes for the fluorescent staining of protein bands in sodium dodecyl sulfate (SDS)-polyacrylamide gels, show an excitation peak in the UV region and another in the visible region (maximum at about 550 nm). Ethidium bromide and other intercalating ... More
A functional virulence complex composed of gingipains, adhesins, and lipopolysaccharide shows high affinity to host cells and matrix proteins and escapes recognition by host immune systems.
AuthorsTakii R, Kadowaki T, Baba A, Tsukuba T, Yamamoto K,
JournalInfect Immun
PubMed ID15664930
'Arg-gingipain (Rgp) and Lys-gingipain (Kgp) are Porphyromonas gingivalis cysteine proteinases implicated as major virulence factors in pathologies of periodontitis. We purified a 660-kDa cell-associated gingipain complex existing as a homodimer of two catalytically active monomers which comprises their catalytic and adhesin domains. Electron microscopy revealed that the complex was composed ... More
Dissociation of steroid receptor coactivator 1 and nuclear receptor corepressor recruitment to the human glucocorticoid receptor by modification of the ligand-receptor interface: the role of tyrosine 735.
AuthorsStevens A, Garside H, Berry A, Waters C, White A, Ray D
JournalMol Endocrinol
PubMed ID12569182
'Within the human glucocorticoid receptor (GR) steroid binding pocket, tyrosine 735 makes hydrophobic contact with the steroid D ring. Substitution of tyrosine735 selectively impairs glucocorticoid transactivation but not transrepression. We now show, using both mammalian two-hybrid and glutathione-S-transferase pull downs, that such substitutions reduce interaction with steroid receptor coactivator 1, ... More
Unusual polypeptide synthesis in the kinetoplast-mitochondria from Leishmania tarentolae. Identification of individual de novo translation products.
AuthorsHorváth A, Nebohácova M, Lukes J, Maslov DA
JournalJ Biol Chem
PubMed ID11773050
'The de novo synthesis of cytochrome c oxidase subunits I, II (COI and COII), and apocytochrome b (Cyb) was investigated in kinetoplast-mitochondria of Leishmania. The organelles were isolated after breaking whole cells with nitrogen cavitation. Individual COI, COII, and Cyb polypeptides were identified by fractionation of the kinetoplast membranes, labeled ... More
Lst1p and Sec24p cooperate in sorting of the plasma membrane ATPase into COPII vesicles in Saccharomyces cerevisiae.
AuthorsShimoni Y, Kurihara T, Ravazzola M, Amherdt M, Orci L, Schekman R
JournalJ Cell Biol
PubMed ID11086000
'Formation of ER-derived protein transport vesicles requires three cytosolic components, a small GTPase, Sar1p, and two heterodimeric complexes, Sec23/24p and Sec13/31p, which comprise the COPII coat. We investigated the role of Lst1p, a Sec24p homologue, in cargo recruitment into COPII vesicles in Saccharomyces cerevisiae. A tagged version of Lst1p was ... More
Determinants for removal and degradation of transit peptides of chloroplast precursor proteins.
AuthorsRichter S, Lamppa GK
JournalJ Biol Chem
PubMed ID12235143
'The stromal processing peptidase (SPP) cleaves a large diversity of chloroplast precursor proteins, removing an N-terminal transit peptide. We predicted previously that this key step of the import pathway is mediated by features of the transit peptide that determine precursor binding and cleavage followed by transit peptide conversion to a ... More
Detection technologies in proteome analysis.
AuthorsPatton WF
JournalJ Chromatogr B Analyt Technol Biomed Life Sci
PubMed ID12015990
'Common strategies employed for general protein detection include organic dye, silver stain, radiolabeling, reverse stain, fluorescent stain, chemiluminescent stain and mass spectrometry-based approaches. Fluorescence-based protein detection methods have recently surpassed conventional technologies such as colloidal Coomassie blue and silver staining in terms of quantitative accuracy, detection sensitivity, and compatibility with ... More
Protein stains for proteomic applications: which, when, why?
AuthorsMiller I, Crawford J, Gianazza E,
JournalProteomics
PubMed ID16991193
'This review recollects literature data on sensitivity and dynamic range for the most commonly used colorimetric and fluorescent dyes for general protein staining, and summarizes procedures for the most common PTM-specific detection methods. It also compiles some important points to be considered in imaging and evaluation. In addition to theoretical ... More
The failure of immune response evasion by linear plasmid 28-1-deficient Borrelia burgdorferi is attributable to persistent expression of an outer surface protein.
AuthorsEmbers ME, Alvarez X, Ooms T, Philipp MT,
JournalInfect Immun
PubMed ID18625742
'Infectivity and persistence by Borrelia burgdorferi, the etiologic agent of Lyme disease, rely stringently on regulatory events. Among these is the downregulation of lipoprotein antigen expression, exemplified by outer surface protein C (OspC), at the advent of specific immunity in the mammalian host. B. burgdorferi spirochetes that lack the linear ... More
Downstream of kinase, p62(dok), is a mediator of Fc gamma IIB inhibition of Fc epsilon RI signaling.
AuthorsOtt VL, Tamir I, Niki M, Pandolfi PP, Cambier JC
JournalJ Immunol
PubMed ID11970986
'The low-affinity receptor for IgG, Fc gamma RIIB, is expressed widely in the immune system and functions to attenuate Ag-induced immune responses. In mast cells, coaggregation of Fc gamma RIIB with the high-affinity IgE receptor, Fc epsilon RI, leads to inhibition of Ag-induced degranulation and cytokine production. Fc gamma RIIB ... More
Chicken histone deacetylase-2 controls the amount of the IgM H-chain at the steps of both transcription of its gene and alternative processing of its pre-mRNA in the DT40 cell line.
AuthorsTakami Y, Kikuchi H, Nakayama T
JournalJ Biol Chem
PubMed ID10446166
'Histone deacetylases (HDACs) are involved in the deacetylation of core histones, which is an important event in transcription regulation in eukaryotes through alterations in the chromatin structure. We cloned cDNAs and genomic DNAs encoding two chicken HDACs (chHDAC-1 and -2), which are preferentially localized in nuclei. Treatment with trichostatin A ... More
Cargo selection into COPII vesicles is driven by the Sec24p subunit.
AuthorsMiller E, Antonny B, Hamamoto S, Schekman R
JournalEMBO J
PubMed ID12426382
'Transport of secretory proteins out of the endoplasmic reticulum (ER) is mediated by vesicles generated by the COPII coat complex. In order to understand how cargo molecules are selected by this cytoplasmic coat, we investigated the functional role of the Sec24p homolog, Lst1p. We show that Lst1p can function as ... More
Proteomics of human breast ductal carcinoma in situ.
AuthorsWulfkuhle JD, Sgroi DC, Krutzsch H, McLean K, McGarvey K, Knowlton M, Chen S, Shu H, Sahin A, Kurek R, Wallwiener D, Merino MJ, Petricoin EF, Zhao Y, Steeg PS
JournalCancer Res
PubMed ID12438275
'We report the first proteomic analysis of matched normal ductal/lobular units and ductal carcinoma in situ (DCIS) of the human breast. An understanding of the transition from normal epithelium to the first definable stage of cancer at the functional level of protein expression is hypothesized to contribute to improved detection, ... More
Sec16p potentiates the action of COPII proteins to bud transport vesicles.
AuthorsSupek F, Madden DT, Hamamoto S, Orci L, Schekman R
JournalJ Cell Biol
PubMed ID12235121
'SEC16 encodes a 240-kD hydrophilic protein that is required for transport vesicle budding from the ER in Saccharomyces cerevisiae. Sec16p is tightly and peripherally bound to ER membranes, hence it is not one of the cytosolic proteins required to reconstitute transport vesicle budding in a cell-free reaction. However, Sec16p is ... More
Coat assembly directs v-SNARE concentration into synthetic COPII vesicles.
AuthorsMatsuoka K, Morimitsu Y, Uchida K, Schekman R
JournalMol Cell
PubMed ID9844642
'COPII proteins are required to create transport vesicles and to select cargo molecules for transit from the ER. A reconstituted liposome budding reaction was used to detect the capture and concentration of membrane-associated v-SNARE molecules into synthetic COPII vesicles. A novel glutathione-phosphatidyl-ethanolamine conjugate (Glut-PE) was synthesized and incorporated into chemically ... More
Fluorescent imaging and analysis with Typhoon 8600.
AuthorsMcNamara P, Lew W, Han L
JournalElectrophoresis
PubMed ID11332750
'Current technologies in laser-based imaging systems for life science applications offer highly sensitive and quantitative detection of multiple labels. Typhoon 8600 variable mode imager is capable of fluorescence, storage phosphor, and chemiluminescence detection of gels and blots. The system delivers sensitive fluorescent detection of nucleic acids and proteins in both ... More
Similar structural basis for membrane localization and protein priming by an RNA-dependent RNA polymerase.
AuthorsLyle JM, Clewell A, Richmond K, Richards OC, Hope DA, Schultz SC, Kirkegaard K
JournalJ Biol Chem
PubMed ID11877407
'Protein primers are used to initiate genomic synthesis of several RNA and DNA viruses, although the structural details of the primer-polymerase interactions are not yet known. Poliovirus polymerase binds with high affinity to the membrane-bound viral protein 3AB but uridylylates only the smaller peptide 3B in vitro. Mutational analysis of ... More
Global role for ClpP-containing proteases in stationary-phase adaptation of Escherichia coli.
AuthorsWeichart D, Querfurth N, Dreger M, Hengge-Aronis R
JournalJ Bacteriol
PubMed ID12486047
'To elucidate the involvement of proteolysis in the regulation of stationary-phase adaptation, the clpA, clpX, and clpP protease mutants of Escherichia coli were subjected to proteome analysis during growth and during carbon starvation. For most of the growth-phase-regulated proteins detected on our gels, the clpA, clpX, or clpP mutant failed ... More
Perspective: proteomics--see "spots" run.
AuthorsKopchick JJ, List EO, Kohn DT, Keidan GM, Qiu L, Okada S
JournalEndocrinology
PubMed ID12021159
Detection of fluorescence dye-labeled proteins in 2-D gels using an Arthur 1442 Multiwavelength Fluoroimager.
AuthorsHerick K, Jackson P, Wersch G, Burkovski A
JournalBiotechniques
PubMed ID11464508
Labeling of proteins with SYPRO Orange, SYPRO Red, and SYPRO Ruby after 2-D polyacrylamide gel electrophoresis (PAGE) using plastic-backed immobilized pH gradient (IPG) strips and precast SDS polyacrylamide gels was tested. Protein spots were detected using an Arthur 1442 Multiwavelength Fluoroimager. The labeling methods described allow detection of proteins both ... More
Determination of total protein content of bacterial cells by SYPRO staining and flow cytometry.
AuthorsZubkov MV, Fuchs BM, Eilers H, Burkill PH, Amann R
JournalAppl Environ Microbiol
PubMed ID10388732
An assay has been developed for measuring protein biomass of marine planktonic bacteria by flow cytometry. The method was calibrated by using five species of Bacteria (an Arcobacter sp., a Cytophaga sp., an Oceanospirillum sp., a Pseudoalteromonas sp., and a Vibrio sp.) recently isolated from seawater samples and grown in ... More
Electrophoretic separation of proteins on a microchip with noncovalent, postcolumn labeling.
AuthorsLiu Y, Foote RS, Jacobson SC, Ramsey RS, Ramsey JM
JournalAnal Chem
PubMed ID11028618
Proteins were separated by microchip capillary electrophoresis and labeled on-chip by postcolumn addition of a fluorogenic dye, NanoOrange, for detection by laser-induced fluorescence. NanoOrange binds noncovalently with hydrophobic protein regions to form highly fluorescent complexes. Kinetic measurements of complex formation on the microchips suggest that the reaction rate is near ... More
Rational design and biophysical characterization of thioredoxin-based aptamers: insights into peptide grafting.
AuthorsBrown CJ, Dastidar SG, See HY, Coomber DW, Ortiz-Lombardía M, Verma C, Lane DP,
JournalJ Mol Biol
PubMed ID19895821
Peptide aptamers are simple structures, often made up of a single-variable peptide loop constrained within a constant scaffold protein. Aptamers were rationally designed by inserting peptides into a solvent-exposed loop on thioredoxin (Trx). They were designed to interact with the proteins elongation initiation factor 4E (eIF4E) and mouse double minute ... More
Cellulase production from spent lignocellulose hydrolysates by recombinant Aspergillus niger.
AuthorsAlriksson B, Rose SH, van Zyl WH, Sjöde A, Nilvebrant NO, Jönsson LJ,
JournalAppl Environ Microbiol
PubMed ID19251882
A recombinant Aspergillus niger strain expressing the Hypocrea jecorina endoglucanase Cel7B was grown on spent hydrolysates (stillage) from sugarcane bagasse and spruce wood. The spent hydrolysates served as excellent growth media for the Cel7B-producing strain, A. niger D15[egI], which displayed higher endoglucanase activities in the spent hydrolysates than in standard ... More
Histometrics: improvement of the dynamic range of fluorescently stained proteins resolved in electrophoretic gels using hyperspectral imaging.
AuthorsWoodward AM, Kaderbhai N, Kaderbhai M, Shaw A, Rowland J, Kell DB,
JournalProteomics
PubMed ID11922594
Most image-based analyses, using absorbance or fluorescence of the spatial distribution of identifiable structures in complex biological systems, use only a very small number of dimensions of possible spectral data for the generation and interpretation of the image. We here extend the concepts of hyperspectral imaging, being developed in remote ... More
Protein gel staining methods: an introduction and overview.
AuthorsSteinberg TH,
JournalMethods Enzymol
PubMed ID19892191
Laboratory scientists who encounter protein biochemistry in many of its myriad forms must often ask: is my protein pure? The most frequent response: run a denaturing SDS polyacrylamide gel. Running this gel raises another series of considerations regarding detection, quantitation, and characterization and so the next questions invariably center on ... More
Multiparameter assessments to determine the effects of sugars and antimicrobials on a polymicrobial oral biofilm.
AuthorsYang Y, Sreenivasan PK, Subramanyam R, Cummins D,
JournalAppl Environ Microbiol
PubMed ID17021225
Clinical studies indicate relationships between dental plaque, a naturally formed biofilm, and oral diseases. The crucial role of nonmicrobial biofilm constituents in maintaining biofilm structure and biofilm-specific attributes, such as resistance to shear and viscoelasticity, is increasingly recognized. Concurrent analyses of the diverse nonmicrobial biofilm components for multiparameter assessments formed ... More
Optimal filter combinations for photographing SYPRO orange or SYPRO red dye-stained gels.
AuthorsSteinberg TH, White HM, Singer VL
JournalAnal Biochem
PubMed ID9177736
Photography or electronic image acquisition is required to document results obtained from staining protein gels with the fluorescent SYPRO dyes. We found that, when using Polaroid type 667 or 57 instant films, the choice of optical filter combination and photographic exposure time strongly influences protein detection sensitivity limits. Ultraviolet light-blocking ... More
Mass spectrometry compatibility of two-dimensional gel protein stains.
As proteomic technology evolves, protein staining sensitivity is constantly being improved, enabling researchers to better visualize the proteome of their system. The current challenge is to balance the limits of detection of protein visualization with those of the mass spectrometric methods. In this report, mass spectra generated from human serum ... More
Applications of SYPRO orange and SYPRO red protein gel stains.
AuthorsSteinberg TH, Haugland RP, Singer VL
JournalAnal Biochem
PubMed ID8811917
We have further characterized the sensitivity and specificity of SYPRO Orange protein gel stain and SYPRO Red protein gel stain with native and 2-dimensional polyacrylamide gels and for staining gels prior to Western blot analysis. We found that nucleic acids are not stained by the SYPRO protein gel stains, in ... More
Subnanomolar detection limit for sodium dodecyl sulfate-capillary gel electrophoresis using a fluorogenic, noncovalent dye.
AuthorsHarvey MD, Bandilla D, Banks PR
JournalElectrophoresis
PubMed ID9761199
Picomolar limits of detection are obtained using the noncovalent, fluorogenic dye, Sypro Red. The size separation of four commonly used sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) molecular weight markers with 8% linear polyacrylamide (PAA) as the sieving matrix is used to construct a calibration curve for molecular weight determinations. SDS-CGE ... More
Postelectrophoretic staining of proteins separated by two-dimensional gel electrophoresis using SYPRO dyes.
AuthorsYan JX, Harry RA, Spibey C, Dunn MJ
JournalElectrophoresis
PubMed ID11271484
While the classical silver stain has been the method of choice for high sensitivity protein visualization on two-dimensional gel electrophoresis (2-D PAGE), post-electrophoretic fluorescent staining with the SYPRO group of dyes has emerged to challenge silver staining for proteome analysis. The latter offers improved sensitivity, higher dynamic range and easy ... More
Yeast Rad17/Mec3/Ddc1: a sliding clamp for the DNA damage checkpoint.
AuthorsMajka J, Burgers PM
JournalProc Natl Acad Sci U S A
PubMed ID12604797
The Saccharomyces cerevisiae Rad24 and Rad17 checkpoint proteins are part of an early response to DNA damage in a signal transduction pathway leading to cell cycle arrest. Rad24 interacts with the four small subunits of replication factor C (RFC) to form the RFC-Rad24 complex. Rad17 forms a complex with Mec3 ... More
Regulation of purified and reconstituted connexin 43 hemichannels by protein kinase C-mediated phosphorylation of Serine 368.
AuthorsBao X, Reuss L, Altenberg GA
JournalJ Biol Chem
PubMed ID14973142
Indirect evidence suggests that the permeability of connexin 43 (Cx43) gap-junctional channels (connexons) to small organic molecules (M(r) < 1,000) is decreased by protein kinase C (PKC)-mediated phosphorylation of Ser-368. However, it is currently unknown whether this effect is produced directly by phosphorylation of this residue or whether cytoplasmic regulatory ... More
A thousand points of light: the application of fluorescence detection technologies to two-dimensional gel electrophoresis and proteomics.
AuthorsPatton WF
JournalElectrophoresis
PubMed ID10786886
As proteomics evolves into a high-throughput technology for the study of global protein regulation, new demands are continually being placed upon protein visualization and quantitation methods. Chief among these are increased detection sensitivity, broad linear dynamic range and compatibility with modern methods of microchemical analyses. The limitations of conventional protein ... More
SYPRO orange and SYPRO red protein gel stains: one-step fluorescent staining of denaturing gels for detection of nanogram levels of protein.
AuthorsSteinberg TH, Jones LJ, Haugland RP, Singer VL
JournalAnal Biochem
PubMed ID8811914
We have developed two new fluorescent dyes, SYPRO Orange protein gel stain and SYPRO Red protein gel stain, to detect proteins in electrophoretic gels. Stained protein bands can be excited by ultraviolet light at approximately 300 nm, or at visible wavelengths, with excitation maxima of 472 nm for the Orange ... More
A luminescent ruthenium complex for ultrasensitive detection of proteins immobilized on membrane supports.
SYPRO Ruby protein blot stain provides a sensitive, gentle, fluorescence-based method for detecting proteins on nitrocellulose or polyvinylidene difluoride (PVDF) membranes. SYPRO Ruby dye is a permanent stain composed of ruthenium as part of an organic complex that interacts noncovalently with proteins. Stained proteins can be excited by ultraviolet light ... More
Requirement for ATP by the DNA damage checkpoint clamp loader.
AuthorsMajka J, Chung BY, Burgers PM
JournalJ Biol Chem
PubMed ID15014082
The DNA damage clamp loader replication factor C (RFC-Rad24) consists of the Rad24 protein and the four small Rfc2-5 subunits of RFC. This complex loads the heterotrimeric DNA damage clamp consisting of Rad17, Mec3, and Ddc1 (Rad17/3/1) onto partial duplex DNA in an ATP-dependent manner. Interactions between the clamp loader ... More
A novel view of gel-shifts: analysis of RNA-protein complexes using a two-color fluorescence dye procedure.
AuthorsShcherbakov D, Piendl W
JournalElectrophoresis
PubMed ID17315147
The electrophoretic mobility shift assay (EMSA) is a common technique to identify and analyze RNA-protein interactions, using the altered electrophoretic mobility of RNA and/or protein upon forming an RNA-protein complex. Traditional techniques of visualization of the EMSA results include either prelabeling of RNA before complex formation or specific RNA- or ... More
Bombesin, lysophosphatidic acid, and epidermal growth factor rapidly stimulate focal adhesion kinase phosphorylation at Ser-910: requirement for ERK activation.
AuthorsHunger-Glaser I, Salazar EP, Sinnett-Smith J, Rozengurt E
JournalJ Biol Chem
PubMed ID12692126
A rapid increase in the tyrosine phosphorylation of focal adhesion kinase (FAK) has been extensively documented in cells stimulated by multiple signaling molecules, but virtually nothing is known about the regulation of FAK phosphorylation at serine residues. Stimulation of Swiss 3T3 cells with bombesin promoted a striking increase ( approximately ... More
Automated ultra-thin-layer SDS gel electrophoresis of proteins using noncovalent fluorescent labeling.
AuthorsCsapo Z, Gerstner A, Sasvari-Szekely M, Guttman A
JournalAnal Chem
PubMed ID10857629
Ultra-thin-layer SDS gel electrophoresis in conjunction with automated laser-induced fluorescence detection is a novel and powerful method for the analysis of fluorophore-labeled proteins. The technique described in this paper employs instant, noncovalent fluorophore labeling by the addition of a fluorescent staining dye to the sample proteins either during or immediately ... More
Making blind robots see: the synergy between fluorescent dyes and imaging devices in automated proteomics.
AuthorsPatton WF
JournalBiotechniques
PubMed ID10818702
Proteomics investigations endeavor to provide a global understanding of gene product synthesis rate, degradation rate, functional competence, posttranslational modification, subcellular distribution and physical interactions with other cell components. Protein expression encompasses an enormous dynamic range. Since rare proteins cannot be amplified by any type of PCR method, sensitive detection is ... More
The ligand affinity of proteins measured by isothermal denaturation kinetics.
AuthorsEpps DE, Sarver RW, Rogers JM, Herberg JT, Tomich PK
JournalAnal Biochem
PubMed ID11319816
An isothermal denaturation kinetic method was developed for identifying potential ligands of proteins and measuring their affinity. The method is suitable for finding ligands specific toward proteins of unknown function and for large-scale drug screening. It consists of analyzing the kinetics of isothermal denaturation of the protein-with and without the ... More
Novel procedure for the identification of proteins by mass fingerprinting combining two-dimensional electrophoresis with fluorescent SYPRO red staining.
AuthorsValdes I, Pitarch A, Gil C, Bermúdez A, Llorente M, Nombela C, Méndez E
JournalJ Mass Spectrom
PubMed ID10862118
The fluorescent sensitive SYPRO Red dye was successfully employed to stain proteins in two-dimensional gels for protein identification by peptide mass fingerprinting. Proteins which are not chemically modified during the SYPRO Red staining process are well digested enzymatically in the gel and hence the resulting peptides can be efficiently eluted ... More
Accessibility of introduced cysteines in chemoreceptor transmembrane helices reveals boundaries interior to bracketing charged residues.
AuthorsBoldog T, Hazelbauer GL
JournalProtein Sci
PubMed ID15133159
Two hydrophobic sequences, 24 and 30 residues long, identify the membrane-spanning segments of chemoreceptor Trg from Escherichia coli. As in other related chemoreceptors, these helical sequences are longer than the minimum necessary for an alpha-helix to span the hydrocarbon region of a biological membrane. Thus, the specific positioning of the ... More
SNARE complex zero layer residues are not critical for N-ethylmaleimide-sensitive factor-mediated disassembly.
AuthorsLauer JM, Dalal S, Marz KE, Nonet ML, Hanson PI
JournalJ Biol Chem
PubMed ID16522630
Membrane-anchored SNAREs assemble into SNARE complexes that bring membranes together to promote fusion. SNARE complexes are parallel four-helix bundles stabilized in part by hydrophobic interactions within their core. At the center of SNARE complexes is a distinctive zero layer that consists of one arginine and three glutamines. This zero layer ... More
Structural and kinetic properties of high and low molecular mass phosphoenolpyruvate carboxylase isoforms from the endosperm of developing castor oilseeds.
AuthorsBlonde JD, Plaxton WC
JournalJ Biol Chem
PubMed ID12519778
Phosphoenolpyruvate carboxylase (PEPC) is believed to play an important role in producing malate as a substrate for fatty acid synthesis by leucoplasts of the developing castor oilseed (COS) endosperm. Two kinetically distinct isoforms of COS PEPC were resolved by gel filtration chromatography and purified. PEPC1 is a typical 410-kDa homotetramer ... More
Analysis of signaling pathways using functional proteomics.
AuthorsResing KA
JournalAnn N Y Acad Sci
PubMed ID12438193
Advances in analytical methods for protein analysis by mass spectrometry provide new tools for global analysis of the expressed protein profile of cells (referred to as proteomics). Currently, available methodology samples only part of the proteome. This is sufficient for analysis of signal transduction, because signaling pathways contain enzymes, which ... More
Proteolytic cleavage of ICAM-1 by human neutrophil elastase.
AuthorsChampagne B, Tremblay P, Cantin A, St Pierre Y
JournalJ Immunol
PubMed ID9834131
Human leukocyte elastase (HLE) participates in tissue destruction in a number of inflammatory disorders, including rheumatoid arthritis and cystic fibrosis. Since HLE has been shown to bind to Mac-1, and ICAM-1 plays a key role during the recruitment and the activation of leukocytes at inflamed sites, we investigated the capacity ... More
Fluorescence detection of proteins in sodium dodecyl sulfate-polyacrylamide gels using environmentally benign, nonfixative, saline solution.
SYPRO Tangerine stain is an environmentally benign alternative to conventional protein stains that does not require solvents such as methanol or acetic acid for effective protein visualization. Instead, proteins can be stained in a wide range of buffers, including phosphate-buffered saline or simply 150 mM NaCl using an easy, one-step ... More
Rapid analysis of covalently and non-covalently fluorophore-labeled proteins using ultra-thin-layer sodium dodecylsulfate gel electrophoresis.
AuthorsGuttman A, Rónai Z, Csapó Z, Gerstner A, Sasvári-Székely M
JournalJ Chromatogr A
PubMed ID11100876
Gel electrophoresis is one of the most frequently used tools for the separation of complex biopolymer mixtures. In recent years, there has been considerable activity in the separation and characterization of protein molecules by sodium dodecylsulfate (SDS) gel electrophoresis with particular interest in using this technique to separate on the ... More
Rpe65 is a retinyl ester binding protein that presents insoluble substrate to the isomerase in retinal pigment epithelial cells.
AuthorsMata NL, Moghrabi WN, Lee JS, Bui TV, Radu RA, Horwitz J, Travis GH
JournalJ Biol Chem
PubMed ID14532273
Photon capture by a rhodopsin pigment molecule induces 11-cis to all-trans isomerization of its retinaldehyde chromophore. To restore light sensitivity, the all-trans-retinaldehyde must be chemically re-isomerized by an enzyme pathway called the visual cycle. Rpe65, an abundant protein in retinal pigment epithelial (RPE) cells and a homolog of beta-carotene dioxygenase, ... More
Are readily culturable bacteria in coastal North Sea waters suppressed by selective grazing mortality?
AuthorsBeardsley C, Pernthaler J, Wosniok W, Amann R
JournalAppl Environ Microbiol
PubMed ID12732530
We studied the growth of six culturable bacterial lineages from coastal North Sea picoplankton in environmental samples under different incubation conditions. The grazing pressure of heterotrophic nanoflagellates (HNF) was reduced either by double prefiltration through 0.8- micro m-pore-size filters or by 10-fold dilutions with 0.2- micro m (pore-size) prefiltered seawater. ... More
Heterogeneous nuclear ribonucleoprotein A1 interferes with the binding of the human T cell leukemia virus type 1 rex regulatory protein to its response element.
AuthorsDodon MD, Hamaia S, Martin J, Gazzolo L
JournalJ Biol Chem
PubMed ID11893730
The human T cell leukemia virus, type 1 (HTLV-1), Rex protein mediates the nuclear export of unspliced and incompletely spliced viral mRNAs. This post-transcriptional activity is dependent in part on the binding of this protein to cis-regulatory sequences termed the Rex-response element (XRE). We have proposed previously that the decreased ... More
Tumor cell killing enabled by listeriolysin O-liposome-mediated delivery of the protein toxin gelonin.
AuthorsProvoda CJ, Stier EM, Lee KD
JournalJ Biol Chem
PubMed ID12832408
Gelonin is a type I plant toxin that has potential as an effective anti-tumor agent by virtue of its enzymatic capacity to inactivate ribosomes and arrest protein synthesis, thereby effectively limiting the growth of cancer cells. Being a hydrophilic macromolecule, however, gelonin has limited access to its target subcellular compartment, ... More
Integration of isoelectric focusing with parallel sodium dodecyl sulfate gel electrophoresis for multidimensional protein separations in a plastic microfluidic [correction of microfludic] network.
AuthorsLi Y, Buch JS, Rosenberger F, DeVoe DL, Lee CS
JournalAnal Chem
PubMed ID14750871
An integrated protein concentration/separation system, combining non-native isoelectric focusing (IEF) with sodium dodecyl sulfate (SDS) gel electrophoresis on a polymer microfluidic chip, is reported. The system provides significant analyte concentration and extremely high resolving power for separated protein mixtures. The ability to introduce and isolate multiple separation media in a ... More