Predictions of protein backbone bond distances and angles from first principles

A procedure is described, based on a spline-function representation of ab initio peptide conformational geometry maps, that allows one to predict backbone bond distances and angles of proteins as functions of the peptide ?(N-C^α)/Ψ(C^α-C′) torsions with an accuracy comparable to that of high-resolution protein crystallography. For example, for the more than 40 residues of crambin, the rms deviation between predicted and crystallographic values of N-C^α-C′ is 1.9° for the 1.5 Å resolution structure and 1.9° for the 0.83 Å resolution structure, compared with angle variations of < 10°. Accurate information on protein backbone geometries is important for establishing dictionaries of flexible geometry functions for use in empirical peptide and protein modeling. © 1995 John Wiley & Sons, Inc.     

Control of the Lysine Biosynthesis Sequence in Corynebacterium glutamicum as Analyzed by Overexpression of the Individual Corresponding Genes

The gene cluster that codes for feedback-resistant aspartate kinase (lysCα and lysCβ) and aspartate semialdehyde dehydrogenase (asd) was cloned from a mutant strain of Corynebacterium glutamicum. Its functional analysis by subcloning, enzyme assays, and type of aspartate kinase regulation enabled the isolation of a fragment for separate expression of the feedback-resistant kinase without aspartate semialdehyde dehydrogenase expression. This was used together with other clones constructed (J. Cremer, L. Eggeling, and H. Sahm, Mol. Gen. Genet. 220:478-480, 1990) to overexpress individually each of the six genes that convert aspartate to lysine. Analysis of lysine formation revealed that overexpression of the feedback-resistant kinase alone suffices to achieve lysine formation (38 mM). Also, sole overexpression of wild-type dihydrodipicolinate synthase resulted in lysine formation but in a lower amount (11 mM). The other four enzymes had no effect on lysine secretion. With a plasmid overexpressing both relevant enzymes together, a further increase in lysine yield was obtained. This shows that of the six enzymes that convert aspartate to lysine the kinase and the synthase are responsible for flow control in the wild-type background and can be useful for construction of lysine-producing strains.     

Zur Kenntnis der OscillatoriaceePseudanabaena galeata (Cyanophyta)

If exsiccated trichomes ofPseudanabaena galeata are immersed in water, striking changes occur as a consequence of lysis and maceration. The locomotion of living trichomes differs from that inOscillatoriaceae. This and other differences make it doubtful, whetherPs. galeata belongs to theOscillatoriaceae; at any rate it occupies an aberrant position.

     

Degradation of coniferyl alcohol and other lignin-related aromatic compounds by Nocardia sp. DSM 1069

Nocardia sp. DSM 1069 was grown on mineral salt media with coniferyl alcohol, 4-methoxybenzoic acid, 3-methoxybenzoic acid or veratric acid as sole sources of carbon and energy. During incubation on coniferyl alcohol, the formation of coniferyl aldehyde, ferulic acid and quantitative accumulation of vanillic acid and proteocatechuic acid could be achieved with mutants. Washed cell suspensions of N. sp. grown on 4-methoxybenzoic acid, oxidized 4-hydroxybenzoic acid and protocatechuic acid. Cells grown on veratric acid, oxidized vanillic acid, isovanillic acid, and protocatechuic acid. Cell extracts were shown to cleave protocatechuic acid by ortho-fission.A mutant without protocatechuate 3,4-dioxygenase activity was not influenced in its growth on 3 methoxybenzoic acid. Cell free extracts of cells grown on 3-methoxybenzoic acid were shown to catalyze the oxidation of gentisic acid (2,5-dihydroxybenzoic acid). The resulting ring cleavage product was further metabolized by a glutathione dependent reaction.The specificity of the demethylation reactions has been investigated with a mutant unable to grow on vanillic acid. This mutant was not impaired in the degradation of isovanillic acid, 4-methoxy-, or 3-methoxybenzoic acid, whereas growth of this mutant on veratric acid (3,4-dimethoxybenzoic acid) was only half as much as that of the wild type. Concomitantly with growth on veratric acid this mutant accumulated vanillic acid with a yield of about 50%.A pathway for the catabolism of coniferyl alcohol, involving oxidation and shortening of the side chain, and of 4-methoxybenzoic acid and veratric acid with protocatechuic acid as intermediate is being proposed. A second one is proposed for the degradation of 3-methoxybenzoic acid with gentisic acid as intermediate.     

Protoprimulagenin a Als Aglykon des Hauptsaponins von Primula elatior

It was shown that the aglycone of the main saponin of Primula elatior (L.) Hill is not primulagenin A but protoprimulagenin A with an oxide ring between C-13 β and C-28 in the β amyrin ring system.     

Lebendbeobachtung der Chromatophorenteilung der DiatomeeNitzschia

Continuous intra vitam observations inNitzschia palea and supplementary ones in two otherN.-species show that the division of the platelike chromatophore is intimately connected with mitosis and cytokinesis. It starts and goes on in a strictly regular course. When the ingrowing plasmatic zone of cytokinetic separation reaches the distal edge of each chromatophore, a narrow split originates and deepens until it has reached the middle of the chromatophore or a little more. Only then a second split arises at the proximal edge of the chromatophore; from there it grows in inverse direction, until it meets the first formed split in a distance of about one third from the proximal end of the chromatophore. Under normal circumstances the position of the meeting point is nearly invariable, the speed of the two progressing splits is the same, and the splitting of the two chromatophores of one cell is nearly exactly synchronized. There is only little variation in the beginning, as the split can form in somewhat different ways. Thereby a striking plasticity of the chromatophore is apparent. The cytokinesis including chromatophore division lasts about 4 1/2–5 minutes. Until now the mode of chromatophore division inNitzschia remains without analogy.     

Atmungsdifferenzen nach Röntgenbestrahlung von Karyopsen bei unterschiedlich strahlensensiblen Weizensorten

An ruhenden und keimenden Karyopsen der Winterweizensorten “Bayro” und “Walthari” wurde die Wirkung einer Röntgenbestrahlung auf die Atmungsintensität untersucht. Bei Bestrahlung lufttrockener Karyopsen liegt die Atmungsintensität von “Bayro” unabhängig von der Höhe der applizierten Dosis wie bei unbestrahlten Karyopsen stets über dem entsprechenden Wert bei “Walthari”. Auch bei Bestrahlung der Karyopsen beider Sorten nach gleichlanger Einquellzeit weist “Bayro” die stärkere Atmung gegenüber “Walthari” auf. Bei Bestrahlung der gequollenen Karyopsen zum Zeitpunkt gleicher Atmungsintensität ergeben sich umgekehrte Verhältnisse. Schon eine Stunde nach der Bestrahlung liegt die Atmungsintensität von “Walthari” über der von “Bayro” und ist auch am Ende der dreistündigen Meßzeit noch mehr als 30% höher. Es werden Möglichkeiten für die Wirkungsweise von Röntgenstrahlen auf die Atmung und der Einfluß unterschiedlicher Strahlensensibilität diskutiert. Frau Marion Kielbasa danke ich für sorgfältige technische Assistenz.