Showing papers in "Methods in 1994"

TL;DR: The immobilization of one binding partner, the possible depletion of analyte close to the sensor surface, and the assumed interaction model are important in the interpretation of binding data.

TL;DR: A rapid, non-radioactive approach to the diagnosis of sickle cell anemia is described based on an allele specific polymerase chain reaction (ASPCR) in which the 3'-terminal nucleotide of one of the primers of the primer set forms a match with one allele and a mismatch with the other allele.

TL;DR: The results obtained indicate that chemically different types of cellulose-bound combinatorial libraries can be prepared easily, allowing the rapid and inexpensive screening of millions of peptides for selection of single molecules with predefined specificity that bind to given ligands such as proteins, metals, nucleic acids, and other molecules of biological interest.

TL;DR: The Ca 2+ -pumping ATPase of the plasma membrane of human erythrocytes can be solubilized with nonionic detergents and purified to homogeneity using calmodulin-affinity chromatography and can be reactivated by the addition of exogenous phospholipids.

TL;DR: Protocols for peptide preparation and handling, for improving instrument sensitivity by background reduction and by HPLC optimizations, and for continued performance assurance are described.

TL;DR: The concept of steric cooperativity is introduced to explain the observed increasing rate of binding as the surface binds more SSB protein and permits calculation of a thermodynamic binding constant.

TL;DR: A functional approach to molecular identification of the large conductance mechanosensitive channel (MscL) from Escherichia coli cell envelope is described and a direct relationship between the mscL product and the channel activity is confirmed by heterologous expression of this gene in two different systems.

TL;DR: In this article, a comprehensive discussion of manual spectrum interpretation is presented, including studies of tryptic, neutral, N-terminally blocked, glycosylated, and other side chain modified peptides.

TL;DR: Methods are described for efficient electrotransfer of proteins onto high-retention PVDF membranes and for N-terminal sequence analysis of electroblotted proteins using either a gas-phase sequencer or a biphasic cartridge sequencer with sequencer cycle times of approximately 40 min.

TL;DR: BIAcore measurements of HIV- 1 serum binding reactivity to peptides corresponding to the third hypervariable (V3) region of gp120 serologically distinguish the two predominate HIV-1 genotypes in Thailand, indicating that the ability of serotyping is also predictive of immunotype.

TL;DR: It is demonstrated that capillary HPLC is an effective method for separating and collecting peptides from one- and two-dimensional polyacrylamide gels, in quantities sufficient for obtaining amino acid sequence data using either Edman methods or mass spectrometry.

TL;DR: A method for in situ digestion that allows high recovery of peptides at the low picomole level from two-dimensional gel electroblotted proteins is described, which provided information sufficient to identify proteins from a protein sequence database using a fragment ion searching algorithm.

TL;DR: The one-bead one-structure process is invaluable in drug discovery for lead identification as well as further optimization of the initial leads and serves as an important research tool for molecular recognition.

TL;DR: In this article, the composition and purity of synthetic combinatorial libraries of free peptides are determined using electrospray mass spectrometry with a maximum of reliability, accuracy, and fastness.

TL;DR: In this paper, a method for multiple column peptide synthesis of resin-bound fluorogenic protease substrates, which are subsequently used in a solid-phase assay for the complete subsite mapping of the active site of endoproteases, is described.

TL;DR: It is found that this "uncaging" of calcium, but not magnesium, barium, or strontium from DM-nitrophen triggers exocytosis, and this threshold appears to be independent of the rate of calcium uncaging.

TL;DR: Initial experiments in this direction with isolated mouse sperm plasma membranes in planar bilayers have led to the detection of a Ca2+ channel resembling one from Strongylocentrotus purpuratus sperm, which suggests that it is important in sperm function.

TL;DR: This article focuses on methods for the use of Xenopus oocytes as a heterologous expression system for plant transporters, and focuses mainly on special considerations applicable to the study of plant iontransporters.

TL;DR: An extremely important application of the BIAcore technology is the characterization of transient, low-affinity interactions, which is used to investigate T-cell receptor (TCR) binding to peptide/MHC molecules.

TL;DR: A simple geometric model is presented as a guide for interpreting interactions among antibodies on an antigen surface and practical guidelines for data reduction and analysis are discussed as well as possible causes of quantitative ambiguity.

TL;DR: A method for the synthesis and screening of polymeric libraries for biologic activities in solution is described, using a construct capable of intramolecular closure of the diketopiperazine ring under neutral conditions in the first stage release.

TL;DR: The use of BlAcore is described to determine the binding constants and specificity of different forms of a recombinant collagen adhesin that all contain the active site but vary in amount of flanking sequence and compare the ligand-binding kinetics of recombinant active sites of four fibronectin-binding adhesins from different bacteria.

TL;DR: In this paper, an interface between liquid chromatography and mass spectrometry is most easily accomplished using electrospray ionization, which is very useful for studying the interactions of proteins with substrates and inhibitors.

TL;DR: Results with two systems, a humanized antibody binding to antigen and coagulation factor VII binding to the cofactor protein tissue factor, are used to describe the use of biosensor technology in mapping protein-protein interfaces and show that the BlAcore instrument is a powerful tool for evaluating protein- protein interactions.

TL;DR: The integration of automated column-based protein chemical modification with automated column's N-terminal protein sequence analysis provides a powerful methodology for obtaining extensive protein structural information without the risks and complications of intervening tedious and costly sample handling steps.

TL;DR: The utility of soluble peptide libraries for the study of antibody/antigen interactions, the identification of highly active opioid peptides in receptor binding studies using crude rat brain homogenates, and in vivo studies, in which the peptide mixtures making up the library are administered intravenously to determine peptide sequences that affect heart rate and blood pressure are illustrated.

TL;DR: The BIAcore has been used successfully to map the location of epitopes in viral antigens and several examples of such studies are presented and it should be possible to establish whether there is a relationship between affinity and neutralizing capacity of antibodies.

TL;DR: Amino acid sequence analysis of peptides, at the picomole level, by triple-quadrupole mass spectrometry is described in this article, where low energy collision-activated dissociation spectra are used to directly search protein and gene sequence databases.