Erratic deposition of agrin during the formation of Xenopus neuromuscular junctions in culture.
AuthorsAnderson MJ, Shi ZQ, Grawel R, Zackson SL
JournalDev Biol
PubMed ID7601300
'In order to disclose the mechanism that regulate synapse development we compared the distributions of agrin, acetylcholine receptors (AChR) and a basal lamina heparan sulfate proteoglycan (HSPG) in sections and cultures prepared from Xenopus laevis and Ambystoma mexicanum embryos. While agrin, AChR and HSPG may accumulate almost synchronously at synapses ... More
A nonradioactive DNA methyltransferase assay adaptable to high-throughput screening.
AuthorsWoo YH, Rajagopalan PT, Benkovic SJ
JournalAnal Biochem
PubMed ID15840507
'We have developed a nonradioactive assay method for DNA methyltransferases based on the ability to protect substrate DNA from restriction. DNA immobilized to a microplate well was treated sequentially with methyltransferase and an appropriate endonuclease. The amount of methylated DNA product is reflected by a proportional decrease in endonuclease cleavage, ... More
Characterization of protein-hapten conjugates. 2. Electrospray mass spectrometry of bovine serum albumin-hapten conjugates.
AuthorsAdamczyk M, Gebler JC, Mattingly PG
JournalBioconjug Chem
PubMed ID8853461
Fifteen hapten-bovine serum albumin (BSA) conjugates were prepared from five commercially available activated haptens. Each hapten was coupled to BSA at three different ratios. The conjugates were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two mass spectrometry (MS) methods: matrix-assisted laser desorption ionization (MALDI) and liquid chromatography-electrospray ... More
Custom fluorescent-nucleotide synthesis as an alternative method for nucleic acid labeling.
AuthorsHenegariu O, Bray-Ward P, Ward DC
JournalNat Biotechnol
PubMed ID10700155
The variety of potentially useful dyes or haptenes available for fluorescent nucleic acid hybridization assays is far greater than what can be obtained from commercial sources. Since this diversity could be useful in many laboratory applications, we have developed a simple and inexpensive procedure for preparing nonpurified labeled nucleotides, for ... More
Mycobacterium avium- and Mycobacterium tuberculosis-containing vacuoles are dynamic, fusion-competent vesicles that are accessible to glycosphingolipids from the host cell plasmalemma.
AuthorsRussell DG, Dant J, Sturgill-Koszycki S
JournalJ Immunol
PubMed ID8648123
The vacuoles inhabited by viable Mycobacterium avium and Mycobacterium tuberculosis show limited fusion with endosomal and lysosomal compartments. This ability to regulate the maturation of their phagosomal compartments and restrict their differentiation into hydrolytically active vacuoles appears to correlate with the survival of the bacilli. Data presented in this current ... More
Cytokine activation leads to acidification and increases maturation of Mycobacterium avium-containing phagosomes in murine macrophages.
AuthorsSchaible UE, Sturgill-Koszycki S, Schlesinger PH, Russell DG
JournalJ Immunol
PubMed ID9570546
Mycobacterium avium (MAC) organisms multiply in phagosomes that have restricted fusigenicity with lysosomes, do not acidify due to a paucity of vacuolar proton-ATPases, yet remain accessible to recycling endosomes. During the course of mycobacterial infections, IFN-gamma-mediated activation of host and bystander macrophages is a key mechanism in the regulation of ... More
Profiling bladder cancer using targeted antibody arrays.
AuthorsSanchez-Carbayo M, Socci ND, Lozano JJ, Haab BB, Cordon-Cardo C
JournalAm J Pathol
PubMed ID16400012
Bladder cancer is a common malignancy requiring a high degree of surveillance because of the frequent recurrences and the poor clinical outcome of invasive disease. To date, serum biomarkers for bladder cancer lack optimal sensitivity and specificity to assist in diagnosis and disease categorization. Here, we designed antibody arrays for ... More
Affinity labeling of a sulfhydryl group in the cardiacglycoside receptor site of Na+/K(+)-ATPase by N-hydroxysuccinimidyl derivatives of digoxigenin.
AuthorsAntolovic R, Linder D, Hahnen J, Schoner W
JournalEur J Biochem
PubMed ID7851443
Na+/K(+)-ATPase from pig kidney is inactivated by protein-reactive N-hydroxysuccinimidyl derivatives of digoxigenin. Like digoxigenin, its protein-reactive derivatives N-hydroxysuccinimidyl digoxigenin-3-methylcarbonyl-epsilon-aminocaproate (HDMA), 3-amino-3-deoxydigoxigenin hemisuccinimide succinimidyl ester (ADHS), 3-iodoacetylamino-3-deoxydigoxigenin (IAD) and digoxigenin-3-O-succinyl-[2-(N-maleimido)]ethylamide (DSME) inhibited the sodium pump in the presence of Na+, Mg2+ and ATP. At 37 degrees C, half-maximal inhibition of Na+/K(+)-ATPase ... More