Voltage-dependent channel formation by rods of helical polypeptides

Summary The voltage-dependence of channel formation by alamethicin and its natural analogues can be described by a dipole flip-flop gating model, based on electric field-induced transbilayer orientational movements of single molecules. These field-induced changes in orientation result from the large permanent dipole moment of alamethicin, which adopts -helical conformation in hydrophobic medium. It was, therefore, supposed that the only structural requirement for voltage-dependent formation of alamethicin-type channels might be a rigid lipophilic helical segment of minimum length.In order to test this hypothesis we synthesized a family of lipophilic polypeptides—Boc-(Ala-Aib-Ala-Aib-Ala) _n -OMe,n=1–4—which adopt -helical conformation forn=2–4 and studied their interaction with planar lipid bilayers. Surprisingly, despite their large difference in chain length, all four polypeptides showed qualitatively similar behavior. At low field strength of the membrane electric field these polypeptides induce a significant, almost voltage-independent increase of the bilayer conductivity. At high field strength, however, a strongly voltage-dependent conductance increase occurs similar to that observed with alamethicin. It results from the opening of a multitude of ion translocating channels within the membrane phase.The steady-state voltage-dependent conductance depends on the 8^th–9^th power of polypeptide concentration and involves the transfer of 4–5 formal elementary charges. From the power dependences on polypeptide concentration and applied voltage of the time constants in voltage-jump current-relaxation experiments, it is concluded that channels could be formed from preexisting dodecamer aggregates by the simultaneous reorientation of six formal elementary charges. Channels exhibit large conductance values of several nS, which become larger towards shorter polypeptide chain length. A mean channel diameter of 19 Å is estimated corresponding roughly to the lumen diameter of a barrel comprised of 10 -helical staves. Similar to experiments with the N-terminal Boc-derivative of alamethicin we did not observe the burst sequence of nonintegral conductance steps typical of natural (N-terminal Ac-Aib)-alamethicin. Saturation in current/voltage curves as well as current inactivation in voltage-jump current-relaxation experiments are found. This may be understood by assuming that channels are generated as dodecamers but, while reaching the steady state, reduce their size to that of an octamer or nonamer. We conclude that the overall behavior of these synthetic polypeptides is very similar to that of alamethicin. They exhibit the same concentration and voltage-dependences but lack the stabilizing principle of resolved channel states characteristic of alamethicin.     

Retinotopische Beziehungen und Struktur rezeptiver Felder im Tectum opticum und Praetectum der Katze

Zusammenfassung 1.Von 600 Neuronen des Colliculus superior und Praetectums der Katze wurde mit Stahlmikroelektroden abgeleitet und der Ableitort markiert. Die Lage der rezeptiven Felder wurde mit bewegten und stationären Lichtreizen bestimmt und dem Ableitort zugeordnet.2.Im Colliculus superior und Praetectum fanden sich richtungsspezifische und richtungsunspezifische Bewegungsneurone. Ein Teil der praetectalen Neurone reagierte richtungsspezifisch auf Bewegungen vom Tier weg und auf das Tier zu (S-Neurone).3.Innerhalb einer senkrecht zur Oberfläche des Colliculus verlaufenden Penetrationssäule nahm die Feldgröße bei gleichbleibender Feldposition mit zunehmender Tiefe zu. Zwischen Ableitort und Feldposition bestand eine systematische retinotopische Beziehung. Die Projektion des vertikalen O-Meridians des Gesichtsfeldes verlief im rostralen Drittel des Colliculus von medial nach lateral, die des horizontalen O-Meridians in der Mitte des Colliculus von rostral nach caudal. Das Projektionsschema eines Colliculus enthält einen nasalen Teil der ipsilateralen Gesichtsfeldhälfte.4.Im Praetectum verlief die Projektion vertikaler Meridiane am caudalen Ende von medial nach lateral und überlappte sich teilweise mit dem Projektionsgebiet des vertikalen O-Meridians im Colliculus. Die horizontalen Meridiane verliefen so von caudal nach rostral, daß das Projektionsschema des Praetectums spiegelbildlich zu dem des Colliculus superior angeordnet war. Dieses Projektionsschema galt nur für den Nucleus tractus optici und die Area praetectalis. Die übrigen praetectalen Kerne mit zum Teil sehr großen rezeptiven Feldern und spezifischen Reaktionsweisen erhielten keine retinotopische Projektion.5.Rezeptive Felder der oberflächennahen Schichten waren uniform on-, off-oder on-off strukturiert, Felder tiefergelegener Einheiten waren ungeordnet aus on-, off- und on-off Bezirken zusammengesetzt. Binocular erregbare Neurone zeigten für beide Augen gleiche Position und Struktur der rezeptiven Felder.6.Die Ergebnisse wurden mit den an anderen Tierarten erhobenen Befunden verglichen. Ihre mögliche funktionelle Bedeutung wurde diskutiert.

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Retinotopic relationship and structure of receptive fields in the optic tectum and pretectum of the cat

Summary 1.600 neurons of the cat's superior colliculus and pretectum were recorded and marked with stainless-steel microelectrodes. The position and structure of receptive fields were tested with stationary flickering and moving stimuli. The position of the stimuli in the visual field was determined by the direction of the lamp projecting the light-points because animal and lamp were arranged in a fixed relationship to the screen. The positions of the stimuli were described in a coordinate system based on the horizontal-and vertical zeromeridean of the retina.2.About 55% of tectal neurons are directionally selective and signal mainly movements directed to the periphery of the visual field. Neurons of the pretectum have the same response characteristics as neurons of the superior colliculus but in addition some are selectively responsive to movements towards the animal or away from it (S-neurons).3.Neurons in one functional column (diameter 0.5 mm, length 3.6 mm) perpendicular to the surface of the superior colliculus react to the same position and preferred direction of a moving stimulus. The size, complexity and directional selectivity of the receptive fields increase with the depth of the recorded neurons. The projection of the vertical zero-meridean passes across the rostral part of the colliculus but does not form the rostral border of the superior colliculus. The nasal part of the ipsilateral visual field projects to the most rostral part of the superior colliculus. The projection of the horizontal zeromeridean passes rostro-caudally in a nearly sagittal plane down the middle of the colliculus. Along this projection-line the resolving power is 13°/column in the caudal part and 6°/column in the rostral part of the superior colliculus. The size of the receptive fields increase with their excentricity in the visual field. (Average of field diameters: 26±13°).4.The diameter of receptive fields in the pretectum was 21±11°, except for a few very large fields (70° and larger). Along the medio-lateral axis of the pretectum there was a retinotopic organization identical to that in the colliculus. Along the caudo-rostral axis, the retinotopic organization was the mirror image of that in the colliculus. No retinotopic organization was observed in the so-called deep pretectal nucleus or in the nucleus of the posterior commissure. Neurons of these nuclei may represent more complex levels in the visual pathway.5.The more superficial neurons of the colliculus (0.1–1.8 mm deep) react mainly with uniform on-, off- or on-off responses to stationary flickered stimuli, i.e. their receptive fields (7–20° in diameter) are uniformly on-, off- or on-off. The deeper neurons (2 mm and deeper) have receptive fields (20–40° in diameter) with compound but not antagonistic structure. No receptive fields showed on- or off-inhibition. Binocularly driven neurons have the same position and structure of their receptive fields for both eyes.6.A survey of the literature reveals that all vertebrates so far investigated show small differences in the destination and retinotopic organization of their retinofugal fibre projections and in the types of tectal receptive fields. These differences seem to indicate an adaption to the development of binocular representation of the center of the visual field, of a specialized area of the retina and of a retino-cortical system.

     

Genetic selection and DNA sequences of 4.5S RNA homologs.

A general strategy for cloning the functional homologs of an Escherichia coli gene was used to clone homologs of 4.5S RNA from other bacteria. The genes encoding these homologs were selected by their ability to complement a deletion of the gene for 4.5S RNA. DNA sequences of the regions encoding the homologs were determined. Since this approach does not require that the homologous genes hybridize with probes generated from the E. coli sequence, the sequences of the homologs were not all similar to the sequence of the E. coli gene. Despite the dissimilarity of the primary sequences of some of the homologs, all could be folded to obtain a similar structure.     

AdaptivSearch: A Novel Iterative Algorithm for the Generation of n-dimensional Hypersurfaces Using an Adaptive Numerical Strategy

AdaptivSearch is the first adaptive strategy based algorithm for the rational and economical construction of n-dimensional hypersurfaces. AdaptivSearch works iteratively: At each step it parcels out the definition range into several triangles, evaluates the worst according to a built-in error criterion, and refines the approximation to the unknown function by choosing the barycenter of this partial area as the node to be calculated next. Based upon the error criterion, AdaptivSearch selectively approaches those parts of the hypersurface in which the curvature exhibits the strongest changes. Some examples of AdaptivSearch applications for both analytical functions and chemical model surfaces are given in order to demonstrate the behavior of the algorithm. These show its broad applicability and the usefulness, especially for chemical problems.     

Acid exposure enhances sporulation of certain strains of Clostridium perfringens

A gastroenteritis results when Clostridium perfringens is ingested in high numbers and sporulates releasing enterotoxin in the intestines. Since the organism must pass through the stomach, its ability to form spores may be affected by the acidic environment. Five strains of C. perfringens were exposed to acidic conditions and then assessed for survival and their ability to form spores. An acidic pH environment kills the bacteria over time but surviving cells are able to recover and form spores. Two of the five strains demonstrated enhanced sporulation following a 30-min exposure to a pH 2 environment. For four of the strains tested, enterotoxin concentrations were higher from acid-exposed cells than from untreated cells. Exposure to a pH 3.5 environment did not affect sporulation when compared to an untreated control. Bacteria in the stationary phase of growth were the most able to resist the acid and sporulate. The results indicate that some strains will produce more spores and enterotoxin following exposure to an acidic environment.     

Directionality of Fission Yeast Mating-Type Interconversion Is Controlled by the Location of the Donor Loci

Cells of homothallic strains of Schizosaccharomyces pombe efficiently switch between two mating types called P and M. The phenotypic switches are due to conversion of the expressed mating-type locus (mat1) by two closely linked silent loci, mat2-P and mat3-M, that contain unexpressed information for the P and M mating types, respectively. In this process, switching-competent cells switch to the opposite mating type in 72-90% of the cell divisions. Hence, mat2-P is a preferred donor of information to mat1 in M cells, whereas mat3-M is a preferred donor in P cells. We investigated the reason for the donor preference by constructing a strain in which the genetic contents of the donor loci were swapped. We found that switching to the opposite mating type was very inefficient in that strain. This shows that the location of the silent cassettes in the chromosome, rather than their content, is the deciding factor for recognition of the donor for each cell type. We propose a model in which switching is achieved by regulating accessibility of the donor loci, perhaps by changing the chromatin structure in the mating-type region, thus promoting an intrachromosomal folding of mat2 or mat3 onto mat1 in a cell type-specific fashion. We also present evidence for the involvement of the Swi6 and Swi6-mod trans-acting factors in the donor-choice mechanism. We suggest that these factors participate in forming the proposed folded structure.     

Der Einfluß von Objektart und früher Erfahrung auf die Diskriminationsleistungen von Haushühnern (Gallus domesticus)3

Main subject of the reported investigation is the question in which way the acquisition of a conditional discrimination is modified on the one hand by the kind of objects presented, on the other hand by early experience. 40 newly hatched chickens grew up either with all the wooden eggs or cubes used for training and test, or without these objects, or in a ‘natural’ (enriched) environment. The results prove an influence of the kind of objects — cubes are discriminated twice or three times as fast as wooden eggs — but no effects of the different environments on acquisition (number of trials for reaching learning criterion). Results are discussed under aspects of species-specific constraints on learning.     

Coordinate regulation of glycogen metabolism in the yeast Saccharomyces cerevisiae. Induction of glycogen branching enzyme.

The yeast glycogen branching enzyme (EC 2.4.1.18) is shown to be induced in batch culture simultaneously with the onset of intracellular glycogen accumulation. The branching enzyme structural gene (GLC3) has been cloned. Its predicted amino acid sequence is very similar to procaryotic branching enzymes. Northern analysis indicates that GLC3 mRNA abundance increases in late exponential growth phase coincident with glycogen accumulation. Disruption of the branching enzyme structural gene establishes that branching enzyme activity is an absolute requirement for maximal glycogen synthesis.     

Serologische Untersuchungen zur Gliederung derBrassicaceae

The serological investigations support the opinion ofJanchen (1942) to combine the generaBunias, Isatis, andSisymbrium in the tribeSisymbrieae; Cheiranthus, Erysimum, andMatthiola in the tribeHesperideae; andBrassica, Crambe, Sinapis, andSuccowia in the tribeBrassiceae. They further underline the central position of theSisymbrieae and the isolated position of theHeliophileae. In accordance withEigner (1973) theBrassiceae are placed closer to theSisymbrieae than inJanchen; the same holds for thePringleeae. No serological justification could be found to uniteArabis andBarbarea in the tribeArabideae, andAlyssum andLunaria in theAlysseae. From the antigen-systems used among the representatives ofJanchen's Lepidieae the generaLepidium andNeslia show remarkable correspondence both toCamelina andThlaspi, but not toCochlearia which appears distant fromCamelina andThlaspi also.

Teil 1/Part 1.     

Biliprotein assembly in the disc-shaped phycobilisomes of Rhodella violacea electron microscopical and biochemical analyses of C-phycocyanin and allophycocyanin aggregates

C-phycocyanin and allophycocyanin from the red alga Rhodella violacea were investigated by electron microscopy and biochemical methods using samples taken from the same fractions.The molecular weights of the native biliprotein aggregates C-phycocyanin and allophycocyanin are about 139,000 (140,000) and 130,000 (145,000) as revealed by calibrated gel chromatography, gradient gel electrophoresis and morphological measurements on the basis of an average protein packing density. These molecular weights are direct evidence for a trimeric aggregation form ()₃ of these biliproteins. Independently, their monomers were determined to be about 34,400 (C-phycocyanin) and 33,900 (allophycocyanin).C-phycocyanin and allophycocyanin are ringshaped, six-membered, biliprotein aggregates with dimensions of about 10.2×3.0 nm and 10.0×3.0 nm, respectively. The aggregates are made up of six subunits, 3 and 3, which are assumed to be associated in alternating positions. They are arranged in regular hexagons in C₆ symmetry. Hexameric aggregates ()₆, so far only isolated for C-phycocyanin, originate by face to face association of two trimeric aggregates.