Conformational analysis of a cyclic thymopoietin-analogue by 1H n.m.r. spectroscopy and restrained molecular dynamics simulations

The internuclear distances of the cyclic thymopoietin derivative c[D-Val-Tyr-Arg-Lys-Glu] have been determined using two-dimensional nuclear Overhauser n.m.r. spectroscopy. These distances are used as constraints for a restrained Molecular Dynamics (MD) simulation. The two starting structures used for the calculations consist of a beta and gamma turn for model 1 and two gamma turns for model 2. The rms difference in atomic positions of the two conformations is 0.242 nm. They converge during the restrained MD simulation to the same final structure. The positional rms difference of the time averaged (5-14 ps) conformations is 0.011 nm. The hydrogen bond pattern is similar to that of model 1, but in addition we find three more gamma turns. The vicinal NH-C alpha H couplings agree well with those calculated from the time averaged structures.     

Sequence-specific 1H-n.m.r. assignments and peptide backbone conformation in rat epidermal growth factor.

The solution conformation of rat epidermal growth factor (EGF) has been investigated by proton n.m.r. techniques. Two-dimensional proton n.m.r. experiments have allowed sequential resonance assignments to be made for most protons. On the basis of these assignments, two regions of anti-parallel beta-sheet structure have been derived from the n.m.r. data. A beta-sheet segment running from about V19 to V23 (capital letters refer to amino acids in the single-letter notation) is folded onto a beta-sheet segment running from R28 to N32 and joined by a chain reversal from E24 to D27. A second region involves a beta-turn from V34 to Y37, which starts a short beta-sheet up to G39, followed by a chain reversal up to Q43, which leads to folding of the C-terminal beta-sheet segment, i.e. H44-R45, running antiparallel to the short Y37 beta-sheet segment. The N-terminal segment up to G18 exists in a multiple bend conformation and is folded on to the V29-V23/R28-N32 beta-sheet such that Y10, Y13, Y22 and Y29 are proximal to each other. Structural comparison of rat, murine and human EGFs indicates a number of highly conserved structural features common to at least these species of EGF.     

Rapid acquisition of three-dimensional triple-resonance experiments using pulsed field gradient techniques

Summary A rapid method for recording three-dimensional triple-resonance experiments utilising pulsed field gradient techniques is proposed, and applied to the HNCO experiment. In order to optimise the sensitivity of the method, a short phase cycle is used in conjunction with the pulsed field gradients to select the desired coherence transfer pathway. The method is demonstrated for the HU protein.     

Photo-CIDNP study of the interaction between lac repressor headpiece and lac operator DNA

Lac repressor headpiece (HP) and intact lac repressor have been studied using the photo-CIDNP method. At neutral pH histidine 29, tyrosines 7, 12 and 17 and methionine 1 are polarised. His-29 polarizations are weaker and broader in HP59 than in HP51 indicating that the C-terminal octapeptide in HP59 adopts a conformation that allows an interaction with His-29. The photo-CIDNP spectra of intact lac repressor and HP51 are very similar, showing that the same residues are accessible to the photo-excited flavin. An equimolar mixture of HP51 and a 14 base pair lac operator fragment strongly suppresses the photo-CIDNP effect of tyrosines 7 and 17 and abolishes the His-29 polarizations. The results are compared with earlier photo-CIDNP measurements on a complex of headpiece with poly[d(AT)] and with a model derived from a 2D NMR study on a lac headpiece-operator complex.     

Stereospecific assignments of 1H-nmr methyl lines and conformation of valyl residues in the lac repressor headpiece

Two-dimensional ¹H-nmr methods are described to obtain information on the sidechain conformation of valyl residues of the lac repressor headpiece and to assign the resonances of their methyl groups stereospecifically. The spin–spin coupling constants (J_αβ) between C^αand C^β protons are obtained from two-dimensional correlated spectroscopy experiments. Large values for J_αβ(10–12 Hz) corresponding to trans orientations for these protons (g⁺ conformation) are found for all valyl residues in α-helical segments. For these valyl residues, the distance between one methyl group (γ₁)and the valyl amide proton is much shorter than for the other methyl group, so that stereospecific resonance assignments follow from relative intensities of the corresponding cross peaks in a two-dimensional nuclear Overhauser enhancement spectrum. Thus, streospecific assignments could be made for the methyl groups of Val 9, 20, 23, and 38 (of a total of eight valyl residues).     

Determination of protein structures from nuclear magnetic resonance data using a restrained molecular dynamics approach: the lac repressor DNA binding domain

A procedure is described to determine from NMR data the three-dimensional structure of biomolecules. This procedure combines model building with a restrained Molecular Dynamics algorithm, in which distance information from NOEs is incorporated in the form of pseudo potentials. The method has been applied to the N-terminal DNA-binding domain or "headpiece" (amino acids 1-51) of the lac repressor from E. coli, for which no crystal structure is available. The spatial structure of the headpiece is discussed in terms of known physical and biochemical data and of its DNA binding properties.     

Alterations in cholesterol and fatty acid biosynthesis in rat liver homogenates by aryloxy acids (Short Communication)

2,4-Dichlorophenoxyacetic acid and 2,4,5-trichlorophenoxyacetic acid inhibited the incorporation of [2-(14)C]mevalonate into cholesterol and non-saponifiable lipids. Both compounds inhibited the conversion of [1-(14)C]isopentenyl pyrophosphate into cholesterol and the synthesis of cholesterol and fatty acids from [2-(14)C]acetate. There was no inhibition of the conversion of [1-(14)C]mevalonate into CO(2). At low concentrations (0.5mm) of the compounds there was a stimulation of acetate incorporation into fatty acids.     

Die auswirkungen der körpergrösse von Aphelinus semiflavus howard (HYM., Aphelinidae) auf einige Organe und ihre leistungsfähigkeit

Zusammenfassung Die Körpergröße von Aphelinus semillavus How. variiert in Abhängigkeit von Große und Qualität des Wirtes. Tiere aus dem gesamten Variationsbereich wurden auf größenabhängige Proportionsverschiebungen untersucht.Relative Länge und Breite des Kopfes (bezogen auf die Körperlange) nehmen mit zunehmender Körperlange ab. Die relative Lange des Abdomens wächst etwa um den gleichen Betrag:Es besteht eine negative Korrelation zwischen der relativen Länge des letzten Fühlergliedes, des Trägers wichtiger Sinnesorgane, und der Körperlange. Seine Oberfläche steht in einem etwa konstanten Verhältnis zur Körperlänge. Die Anzahl der Porenplatten nimmt mit der Körperlänge nur wenig, ihre Länge deutlicher zu.Im Komplexauge wächst die Anzahl der Ommatidien etwa proportional zur Korperlange, während seine gesamte Fläche zwar linear, aber ïiberproportional ansteigt, was durch zunehmende Große der einzelnen Facetten erreicht wird.Die Länge der Vorderfliigel ist bei Männchen und Weibchen von mehr als 1 mm Länge etwa der Körperlange proportional. Unterhalb 1 mm nimmt die relative Flügellänge mit der Körperlange ab, bis die Flügel nur noch funktionslose Rudimente bilden. Die Relation Flügel-fläche:Körpergewicht ist bei mittelgroßen Tieren am günstigsten.Jedes Ovar hat stets drei Ovariolen, unabhängig von der Körperlänge. Große Weibchen legen nur etwas größere, aber bedeutend mehr Eier als kleine. Die Anzahl der (am 7.–10. Lebenstag) täglich abgelegten Eier steigt etwa linear mit dem Logarithmus des Körpergewichts an.Die Eiablage wird während des ganzen Lebens (im Labor etwa 40 Tage) fortgesetzt, ist aber zum. Ende hin etwas geringer. Relativ zum Körper-gewicht produzieren mittelgroße Weibchen die größte Eimasse. Die Anzahl der zum Nahrungsgewinn ausgesaugten Läuse steigt mit dem Körpergewicht.Zwischen der relativen Länge des Legestachels und der Körperlange besteht eine fast lineare negative Korrelation.Die gröBenabhangigen Proportionsverschiebungen werden als Auswirkungen unterschiedlicher Wachstumstendenzen der einzelnen Organe gedeutet, die ihrerseits genetisch fixiert sind und den biologischen Sinn haben, die Lebensf ähigkeit modifikatorisch kleiner Exemplare zu sichern und damit die Ausnutzung entsprechend kleiner Wirte zu ermöglichen.Die Bedeutung der Proportionsverschiebungen für die Leistung der ganzen Population wird diskutiert. Sic ist abhängig von der Häufigkeits-verteilung der verschiedenen Körpergrößen und these wiederum von der Art-Zusammensetzung und Ernährung der Wirtsfauna.

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Summary The body size of Aphelinus semiflavus How. varies with the size and the quality of the hosts. Specimens from the whole range of variation were investigated for differences between organ proportions, which depended on size.The relative length and width of the head (based on the body length) are reduced with increasing body length. The relative length of the abdomen increases approximately by the same amount.There is a negative correlation between the relative length of the last joint of the antennae and the body length. Its surface shows a nearly constant proportion to the body length. The length of the pore plates increases with the body length more distinctly than their numbers.Within the compound eye, the number of ommatidia increases almost proportionally to the body length; the whole area of the eye increases more than proportionally because the size of the single facets also becomes greater.When the body length is greater than 1 mm, the length of the forewings is nearly proportional to the body length. At less than 1 mm, the relative length of the fore-wings is reduced with declining body length until, finally, only rudiments without any function remain. The ratio between wing area and body weight is most favourable at medium body size.Three ovarioles, independent of the body size, are present in each ovary. Big females produce considerably more eggs than small ones. The size of their eggs is only slightly enlarged. The daily egg production (at the 7^th–10^th day of life) increases nearly linearly with the logarithm of body weight. Oviposition is continued throughout the life-time of the insect (ca. 40 days in the laboratory) but it is somewhat reduced at the end. Relative to their body weight, females of medium size produce the biggest egg masses. The number of aphids utilized by the parasite for nourishment increases with the body weight of the female.A negative correlation exists between the relative length of the extended ovipositor and the body length.The proportion differences of the organs, dependent on size, are interpreted to be consequences of different growth tendencies which are genetically fixed. Apparently, the biological role of such different growth tendencies is to ensure the viability of phenotypically small specimens and in this way to make the utilisation of small hosts possible.The importance of the differences of body proportions for the effect of the whole population is discussed. It depends on the frequency distribution of the different body lengths which itself is the consequence of the composition of host species and the nutriation of the hosts.

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Herrn Prof. Dr. Hans Braun, emer. Direktor des Instituts für Pfanzenkrankheiten an der Universität Bonn, zu seinem 70. Geburts'ag am 2. Februar 1966 gewidmet.