Restriction of receptor movement alters cellular response: physical force sensing by EphA2.
AuthorsSalaita K, Nair PM, Petit RS, Neve RM, Das D, Gray JW, Groves JT,
JournalScience
PubMed ID20223987
'Activation of the EphA2 receptor tyrosine kinase by ephrin-A1 ligands presented on apposed cell surfaces plays important roles in development and exhibits poorly understood functional alterations in cancer. We reconstituted this intermembrane signaling geometry between live EphA2-expressing human breast cancer cells and supported membranes displaying laterally mobile ephrin-A1. Receptor-ligand binding, ... More
Proinflammatory and vasodilator effects of nociceptin/orphanin FQ in the rat mesenteric microcirculation are mediated by histamine.
'Nociceptin/orphanin FQ (N/OFQ) is the endogenous ligand for the N/OFQ peptide receptor (NOP). N/OFQ causes hypotension and vasodilation, and we aimed to determine the role of histamine in inflammatory microvascular responses to N/OFQ. Male Wistar rats (220-300 g, n = 72) were anesthetized with thiopental (30 mg/kg bolus, 40-90 mg ... More
Green- and red-fluorescent nanospheres for the detection of cell surface receptors by flow cytometry.
AuthorsBhalgat MK, Haugland RP, Pollack JS, Swan S, Haugland RP
JournalJ Immunol Methods
PubMed ID9831388
'Fluorescent probes serve as sensitive tools for obtaining structural and functional information in cellular systems. In spite of the high sensitivity provided by fluorescent reagents, cell surface receptors expressed in low numbers often escape detection with commonly used fluorescent probes. R-Phycoerythrin (R-PE), a molecule with a very high quantum yield, ... More
Micromechanical tests of adhesion dynamics between neutrophils and immobilized ICAM-1.
AuthorsLomakina EB, Waugh RE
JournalBiophys J
PubMed ID14747356
'Strong, integrin-mediated adhesion of neutrophils to endothelium during inflammation is a dynamic process, requiring a conformational change in the integrin molecule to increase its affinity for its endothelial counterreceptors. To avoid general activation of the cell, Mg(2+) was used to induce the high-affinity integrin conformation, and micromechanical methods were used ... More
Reversible self-assembly and directed assembly of DNA-linked micrometer-sized colloids.
AuthorsValignat MP, Theodoly O, Crocker JC, Russel WB, Chaikin PM
JournalProc Natl Acad Sci U S A
PubMed ID15758072
'We present a technique for the directed assembly and self-assembly of micrometer-scale structures based on the control of specific DNA linkages between colloidal particles. The use of DNA links combined with polymer brushes provides an effective way to regulate the range and magnitude of addressable forces between pairs (and further ... More
Recombinant P-selectin glycoprotein ligand-1 directly inhibits leukocyte rolling by all 3 selectins in vivo: complete inhibition of rolling is not required for anti-inflammatory effect.
AuthorsHicks AE, Nolan SL, Ridger VC, Hellewell PG, Norman KE,
JournalBlood
PubMed ID12480716
'Selectin-dependent leukocyte rolling is one of the earliest steps of an acute inflammatory response and, as such, contributes to many inflammatory diseases. Although inhibiting leukocyte rolling with selectin antagonists is a strategy that promises far-reaching clinical benefit, the perceived value of this strategy has been limited by studies using inactive, ... More
The helicase domain and C-terminus of human RecQL4 facilitate replication elongation on DNA templates damaged by ionizing radiation.
AuthorsKohzaki M, Chiourea M, Versini G, Adachi N, Takeda S, Gagos S, Halazonetis TD,
JournalCarcinogenesis
PubMed ID22508716
The vertebrate RECQL4 (RECQ4) gene is thought to be the ortholog of budding yeast SLD2. However, RecQL4 contains within its C-terminus a RecQ-like helicase domain, which is absent in Sld2. We established human pre-B lymphocyte Nalm-6 cells, in which the endogenous RECQL4 gene was homozygously targeted such that the entire ... More
Liquid-based fluorescence in situ hybridization assay for detection of ERBB2 gene amplification in patients with breast cancer.
AuthorsYeh CH, Whitmire WA, Albitar M,
JournalClin Chem
PubMed ID18787015
Current reference methods for evaluating gene amplification and expression of ERBB2 (also known as HER-2)--cell-based fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC)--are subjective and influenced by methods of tissue preparation and fixation. We developed and evaluated a novel, quantitative liquid-based FISH (L-FISH) assay that uses flow cytometry to detect ... More
A novel high-throughput screen reveals yeast genes that increase secretion of heterologous proteins.
AuthorsWentz AE, Shusta EV,
JournalAppl Environ Microbiol
PubMed ID17189442
The yeast Saccharomyces cerevisiae is an attractive host for the production of heterologous proteins. However, low-yield production of many proteins (from micrograms to milligrams/liter) leaves considerable room for optimization. By engineering the yeast cell via traceable genome-wide libraries, genes that can enhance protein expression level because of their roles in ... More
Leukocyte function-associated antigen 1-mediated adhesion stability is dynamically regulated through affinity and valency during bond formation with intercellular adhesion molecule-1.
AuthorsSarantos MR, Raychaudhuri S, Lum AF, Staunton DE, Simon SI,
JournalJ Biol Chem
PubMed ID15955822
Neutrophil rolling and transition to arrest on inflamed endothelium are dynamically regulated by the affinity of the beta(2) integrin CD11a/CD18 (leukocyte function associated antigen 1 (LFA-1)) for binding intercellular adhesion molecule (ICAM)-1. Conformational shifts are thought to regulate molecular affinity and adhesion stability. Also critical to adhesion efficiency is membrane ... More
Identification of peptides targeting the surface of Plasmodium falciparum-infected erythrocytes using a phage display peptide library.
AuthorsEda K, Eda S, Sherman IW,
JournalAm J Trop Med Hyg
PubMed ID15306709
Drastic changes in the plasma membrane of Plasmodium falciparum-infected red blood cells (iRBCs) make the surface of iRBCs distinct from that of the uninfected erythrocyte. To identify small peptides that would specifically recognize the altered surface of iRBCs, we screened a phage display peptide library (PDL) on the surface of ... More
Direct visualization of cystic fibrosis transmembrane regulator mutations in the clinical laboratory setting.
AuthorsStrom CM, Clark DD, Hantash FM, Rea L, Anderson B, Maul D, Huang D, Traul D, Chen Tubman C, Garcia R, Hess PP, Wang H, Crossley B, Woodruff E, Chen R, Killeen M, Sun W, Beer J, Avens H, Polisky B, Jenison RD,
JournalClin Chem
PubMed ID15010427
The recommendation for population- based cystic fibrosis (CF) carrier screening by the American College of Medical Genetics for the 25 most prevalent mutations and 6 polymorphisms in the CF transmembrane regulatory gene has greatly increased clinical laboratory test volumes. We describe the development and technical validation of a DNA chip ... More
Granulocyte colony-stimulating factor promotes adhesion of neutrophils.
AuthorsChakraborty A, Hentzen ER, Seo SM, Smith CW,
JournalAm J Physiol Cell Physiol
PubMed ID12388113
Granulocyte colony stimulating factor (G-CSF) is well known for its ability to drive the maturation and mobilization of neutrophils. G-CSF also appears to have the potential to activate functions of mature neutrophils, influencing recruitment at sites of inflammation and tissue injury. We investigated the ability of G-CSF to stimulate adhesion ... More
Effects of IL-8, Gro-alpha, and LTB(4) on the adhesive kinetics of LFA-1 and Mac-1 on human neutrophils.
AuthorsSeo SM, McIntire LV, Smith CW,
JournalAm J Physiol Cell Physiol
PubMed ID11600420
Firm adhesion of rolling neutrophils on inflamed endothelium is dependent on beta(2) (CD18)-integrins and activating stimuli. LFA-1 (CD11a/CD18) appears to be more important than Mac-1 (CD11b/CD18) in neutrophil emigration at inflammatory sites, but little is known of the relative binding characteristics of these two integrins under conditions thought to regulate ... More
Comparison of detection and signal amplification methods for DNA microarrays.
AuthorsVora GJ, Meador CE, Anderson GP, Taitt CR,
JournalMol Cell Probes
PubMed ID18675897
One of the factors limiting the use of DNA microarray technology for the detection of pathogenic organisms from clinical and environmental matrices has been inadequate assay sensitivity. To assess the effectiveness of post-hybridization secondary detection steps to enhance the sensitivity of DNA microarray-based pathogen detection, we evaluated a panel of ... More
Cross talk between tetanus neurotoxin-insensitive vesicle-associated membrane protein-mediated transport and L1-mediated adhesion.
AuthorsAlberts P, Rudge R, Hinners I, Muzerelle A, Martinez-Arca S, Irinopoulou T, Marthiens V, Tooze S, Rathjen F, Gaspar P, Galli T
JournalMol Biol Cell
PubMed ID14517330
The membrane-trafficking pathway mediated by tetanus neurotoxin-insensitive vesicle-associated membrane protein (TI-VAMP) in neurons is still unknown. We show herein that TI-VAMP expression is necessary for neurite outgrowth in PC12 cells and hippocampal neurons in culture. TI-VAMP interacts with plasma membrane and endosomal target soluble N-ethylmaleimide-sensitive factor attachment protein receptors, suggesting ... More