Elucidation of a PTS–Carbohydrate Chemotactic Signal Pathway in Escherichia coli Using a Time-resolved Behavioral Assay

Chemotaxis of Escherichia coli toward phosphotransferase systems (PTSs)–carbohydrates requires phosphoenolpyruvate-dependent PTSs as well as the chemotaxis response regulator CheY and its kinase, CheA. Responses initiated by flash photorelease of a PTS substrates d-glucose and its nonmetabolizable analog methyl α-d-glucopyranoside were measured with 33-ms time resolution using computer-assisted motion analysis. This, together with chemotactic mutants, has allowed us to map out and characterize the PTS chemotactic signal pathway. The responses were absent in mutants lacking the general PTS enzymes EI or HPr, elevated in PTS transport mutants, retarded in mutants lacking CheZ, a catalyst of CheY autodephosphorylation, and severely reduced in mutants with impaired methyl-accepting chemotaxis protein (MCP) signaling activity. Response kinetics were comparable to those triggered by MCP attractant ligands over most of the response range, the most rapid being 11.7 ± 3.1 s−1. The response threshold was <10 nM for glucose. Responses to methyl α-d-glucopyranoside had a higher threshold, commensurate with a lower PTS affinity, but were otherwise kinetically indistinguishable. These facts provide evidence for a single pathway in which the PTS chemotactic signal is relayed rapidly to MCP–CheW–CheA signaling complexes that effect subsequent amplification and slower CheY dephosphorylation. The high sensitivity indicates that this signal is generated by transport-induced dephosphorylation of the PTS rather than phosphoenolpyruvate consumption.

Structure and function in rhodopsin: Further elucidation of the role of the intradiscal cysteines, Cys-110, -185, and -187, in rhodopsin folding and function

The disulfide bond between Cys-110 and Cys-187 in the intradiscal domain is required for correct folding in vivo and function of mammalian rhodopsin. Misfolding in rhodopsin, characterized by the loss of ability to bind 11-cis-retinal, has been shown to be caused by an intradiscal disulfide bond different from the above native disulfide bond. Further, naturally occurring single mutations of the intradiscal cysteines (C110F, C110Y, and C187Y) are associated with retinitis pigmentosa (RP). To elucidate further the role of every one of the three intradiscal cysteines, mutants containing single-cysteine replacements by alanine residues and the above three RP mutants have been studied. We find that C110A, C110F, and C110Y all form a disulfide bond between C185 and C187 and cause loss of retinal binding. C185A allows the formation of a C110–C187 disulfide bond, with wild-type-like rhodopsin phenotype. C187A forms a disulfide bond between C110 and C185 and binds retinal, and the pigment formed has markedly altered bleaching behavior. However, the opsin from the RP mutant C187Y forms no rhodopsin chromophore.

SURVEY AND SUMMARY: Recent advances in the elucidation of the mechanisms of action of ribozymes

The cleavage of RNA can be accelerated by a number of factors. These factors include an acidic group (Lewis acid) or a basic group that aids in the deprotonation of the attacking nucleophile, in effect enhancing the nucleophilicity of the nucleophile; an acidic group that can neutralize and stabilize the leaving group; and any environment that can stabilize the pentavalent species that is either a transition state or a short-lived intermediate. The catalytic properties of ribozymes are due to factors that are derived from the complicated and specific structure of the ribozyme–substrate complex. It was postulated initially that nature had adopted a rather narrowly defined mechanism for the cleavage of RNA. However, recent findings have clearly demonstrated the diversity of the mechanisms of ribozyme-catalyzed reactions. Such mechanisms include the metal-independent cleavage that occurs in reactions catalyzed by hairpin ribozymes and the general double-metal-ion mechanism of catalysis in reactions catalyzed by the Tetrahymena group I ribozyme. Furthermore, the architecture of the complex between the substrate and the hepatitis delta virus ribozyme allows perturbation of the pKa of ring nitrogens of cytosine and adenine. The resultant perturbed ring nitrogens appear to be directly involved in acid/base catalysis. Moreover, while high concentrations of monovalent metal ions or polyamines can facilitate cleavage by hammerhead ribozymes, divalent metal ions are the most effective acid/base catalysts under physiological conditions.

The natural history of the human teeth including a particular elucidation of the changes which take place during the second dentition and describing the proper mode of treatment to prevent irregularities of the teeth... /

Mode of access: Internet.

Federal taxes for 1923; elucidation of the law, the Revenue law of 1921, the Amendments of March, 1923, tables illustrating taxes; indexed.

Mode of access: Internet.

The sportsman's cyclopaedia : comprising a complete elucidation of the science and practice of hunting, shooting, coursing, racing, fishing, hawking, cockfighting, and other sports and pastimes of Great Britain, interspersed with entertaining and illustrative anecdotes /

Extra numbered pages: 183*-184* and 191*-196* inserted.

Nineveh and its palaces : the discoveries of Botta and Layard, applied to the elucidation of holy writ /

Mode of access: Internet.

An elucidation of the unity of God : deduced from Scripture and reason, addressed to Christians of all denominations : to which is subjoined a letter from the author to his Grace the Archbishop of Canterbury, 3d ed., with additions.

Mode of access: Internet.

An account of a most dangerous voyage peformed by the famous Captain John Monck, in the years 1619, and 1620 : by the special command of Cristian IV : to Hudson's Straits, in order to discover a passage on that side, betwixt Greenland and America to the West [read East]-Indies : with a description of the old and new Greenland, for the better elucidation of the said treatise /

Mode of access: Internet.

Dictionary of the English and German languages for general use : compiled with especial regard to the elucidation of modern literature ... /

Mode of access: Internet.

Animal heroes : being the histories of a cat, a dog, a pigeon, a lynx, two wolves & a reindeer and in elucidation of the same over 200 drawings /

Full page ill. on p. 11, 27, 43, 71, 83, 105, 109, 119, 131, 145, 161, 167, 175, 189, 201, 205, 225, 252, 257, 265, 283, 287, 293, 305, 321, 339 and 363, some signed by Ernest Thompson Seton. Incidental illustrations throughout.

A compilation of genuine church music : comprising a variety of metres, all harmonized for three voices : together with a copious elucidation of the science of vocal music /

Includes indexes.

A critical and practical elucidation of the Book of Common Prayer, and administration of the sacraments, and other rites and ceremonies of the church, according to the use of the United Church of England and Ireland /

Mode of access: Internet.