Gcorge M. Whitesides

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This paper describes the estimation of binding constants (Kb) between carbonic anhydrase B (CAB, EC 4.2.1.1, from bovine erythrocytes) and charged benzenesulfonamides by affinity capillary electrophoresis (ACE) under conditions in which the migration time is affected by changes in electroosmotic flow and by nonspecific interactions accompanying changes in(More)
An alpha-sialoside linked to acrylamide by a short connector (5-acetamido-2-O-(N-acryloyl-8-amino-5-oxaoctyl)-2,6-anhydro-3,5-d ideoxy-D-galacto-alpha-nonulopyranosonoic acid, 1) was prepared. Compound 1 formed high molecular weight copolymers with acrylamide, derivatives of acrylamide, and/or vinylpyrrolidone upon photochemically-initiated free radical(More)
A series of competitive inhibitors of carbonic anhydrase II (CAII; EC 4.2.1.1) that consists of oligo(ethylene glycol) units attached to p-benzenesulfonamides with pendant amino acids, H2NSO2C6H4CONHCH2CH2OCH2CH2OCH2CH2NHCOCHRNH3+, have been synthesized and examined using competitive fluorescence assays. Three of the strongest inhibitors, designated(More)
This paper describes convenient preparations of heterogeneous multisectional polyacrylamide slab gels and the protocols that use these gels to measure protein-ligand binding constants [using bovine carbonic anhydrase B (CAB) as a model system]. Unlike procedures for affinity electrophoresis using tube gels, all binding information concerning protein-ligand(More)
A practical l5-ntmol large-scale synthcsis ol p-r.-fucopy'ranosyl dicyclohex-vlammonium phosphatc frttm t-fucose in 63cl overall yield was dcvclope d. The synthcsis took advantagc of a ncighboring I3z-2 group participating in a Koenigs-Knorr-like glycosvlation. The sugur phosphate was translitrnrecl into the activ:rted sugar nucleoside. guanosine(More)
This paper describes cell-free enzymatic syntheses of sucrose and trehalose using partially-purified preparations of sucrose and trehalose synthetase. The coupling of the regeneration of uridine-5'-diphosphoglucose (UDP-Glc) with synthesis of the disaccharide offers a practical route to millimol quantities of these carbohydrates. The syntheses used pyruvate(More)
Agarose beads containing immobilized enzymes or affinity ligands have been made magnetically responsive by adsorbing freshly precipitated magnetite on their surface. These beads are used for affinity adsorption of proteins from complex mixtures containing suspended solids. The magnetically responsive beads and the unwanted (diamagnetic) solids are then(More)
Cell-free protein synthesis in rabbit reticulocyte lysate translation mixtures has been studied during multi-hour incubations. In an impaired lysate obtained from cells stored at 0 degrees C before lysis, and showing a low initial rate of synthesis, translation could be stimulated by a factor of 4 by including RNase inhibitor and additional ATP and GTP. In(More)
To measure the depth of the local anesthetic binding site within the neuronal membrane, biotin-containing polyethylene glycols having zero, three, and six ethylene glycol subunits were added to the p-amino termini of tetracaine and procaine, thereby interposing a pharmacologically inert "spacer" molecule between the local anesthetic and the biotin moiety.(More)
Affinity capillary electrophoresis (ACE) provides a new approach to studying protein-ligand interactions. The basis for ACE is the change in the electrophoretic mobility of the protein when it forms a complex with its ligand. This binding interaction can be quantified directly for charged ligands or indirectly for neutral ligands in competition with a(More)