Evidence for essential primary amino groups in a bacterial coupling factor F1ATPase

We have found that the binding of pyridoxal-5′-phosphate to 6 primary amino groups leads to the inactivation of the enzyme. A preferential reaction of pyridoxal-5′-phosphate with the α-subunits of this enzyme can be demonstrated. The reactivity of the amino groups is influenced by various effectors. In the presence of ATP the inhibition of the ATPase activity is noncompetitive.     

Automated extraction of bank angles from bird‐borne video footage using open‐source software

The use of miniaturized video cameras to study the at‐sea behavior of flying seabirds has increased in recent years. These cameras allow researchers to record several behaviors that were not previously possible to observe. However, video recorders produce large amounts of data and videos can often be time‐consuming to analyze. We present a new technique using open‐source software to extract bank angles from bird‐borne video footage. Bank angle is a key facet of dynamic soaring, which allows albatrosses and petrels to efficiently search vast areas of ocean for food. Miniaturized video cameras were deployed on 28 Wandering Albatrosses (Diomedea exulans) on Marion Island (one of the two Prince Edward Islands) from 2016 to 2018. The OpenCV library for the Python programming language was used to extract the angle of the horizon relative to the bird’s body (= bank angle) from footage when the birds were flying using a series of steps focused on edge detection. The extracted angles were not significantly different from angles measured manually by three independent observers, thus being a valid method to measure bank angles. Image quality, high wind speeds, and sunlight all influenced the accuracy of angle estimates, but post‐processing eliminated most of these errors. Birds flew most often with cross‐winds (58%) and tailwinds (39%), resulting in skewed distributions of bank angles when birds turned into the wind more often. Higher wind speeds resulted in extreme bank angles (maximum observed was 94°). We present a novel method for measuring postural data from seabirds that can be used to describe the fine‐scale movements of the dynamic‐soaring cycle. Birds appeared to alter their bank angle in response to varying wind conditions to counter wind drift associated with the prevailing westerly winds in the Southern Ocean. These data, in combination with fine‐scale positional data, may lead to new insights into dynamic‐soaring flight.     

A new synthetic approach to 1-[(3R,4R)-1-cyclooctylmethyl-3-hydroxymethyl-4-piperidyl]-3-ethyl-1,3-dihydro-benzimidazol-2-one(J-113397), the first non-peptide ORL-1 receptor antagonist..

An efficient approach to 1-[(3R,4R)-1-cyclooctylmethyl-3-hydroxymethyl-4-piperidyl]-3-ethyl-1,3-dihydro-benzimidazol-2-one (J-113397) 1, the first non-peptide ORL-1 receptor antagonist described in literature, is outlined. After construction of the piperidine framework through Dieckmann cyclization of the Michael adduct 8 of cyclooctylmethylamine to methyl acrylate, condensation with o-phenylendiamine produced the beta-enamino ester 2, which has been conveniently used to construct the benzimidazolone substituent at C-4. Catalytic hydrogenation of intermediate 11 followed by base-promoted cis--trans isomerization of the key compound 12 led to the formation of ester 13, which was converted to the racemic title compound by LiAlH(4) reduction. The pure enantiomers were obtained by chiral preparative HPLC separation using a derivatized cellulose-based stationary phase.     

High-Resolution Cryo-EM Structure of the Cardiac Actomyosin Complex

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N-Terminal Domains of Cardiac Myosin Binding Protein C Cooperatively Activate the Thin Filament

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Veränderung von Aminosäuren und Aktivität bei Röntgenbestrahlung von krystallisiertem Lysozym

Zusammenfassung Kristallisiertes Lysozym wurde mit Dosen zwischen 1 und 100 Mrad Röntgenstrahlen bestrahlt. Die kleineren Dosen haben keine meßbare Wirkung auf Aktivität oder quantitative Aminosäurezusammensetzung. Da das Molekül durch größere Strahlendosen in andere Modifikationen umgewandelt wird und die entsprechenden Vorgänge komplex sind, läßt sich die naktivierungdes Enzyms nicht ohne weiteres durch die lassische Treffertheorie interpretieren. Hiermit zusammenhängende Probleme werden diskutiert. Erst Strahlendosen von der Größenordnung 100 Mrad erzeugen in einem automatischen Aminosäureanalysator sicher analysierbare Aminosäureveränderungen. Die G-Werte liegen zwischen 3 und 13. Die größere Strahlenempfindlichkeit haben vor allem die aromatischen und die schwefelhaltigen Aminosäuren. Vergleiche mit anderen Ergebnissen führen zu dem Schluß, daß diese Strahlenempfindlichkeit der Aminosäuren durch Milieufaktoren nur wenig beeinflußt wird.

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Summary Crystallized lysozyme was X-irradiated with doses between 1 and 100 Mrads. Doses of a few Mrads have no measurable effect on activity and quantitative amino acid composition of lysozyme. However, increasing doses successively convert the original protein into a large number of modified proteins showing more or less lysozyme activity. These processes are of a complex nature. Consequently can the inactivation not be described in terms of the classical target theory. Only X-ray doses of the order of magnitude of 100 Mrads produce amino acid changes which can clearly be determined with an automatic amino acid analyzer. The G-values for the destruction of the various constituent amino acids are between 3 and 13. The larger sensitivities have the S-containing and aromatic amino acids as well as the long chain aliphatic amino acids. A comparison with other results shows that the radiation sensitivity of the constituent amino acids of solid proteins is little influenced by other factors (O₂-pressure, water content, protein structure etc.).

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Auszugsweise vorgetragen während des ESR-Symposiums Gatlinburg, Tenn., Mai 1965, und während eines Chemischen Kolloquiums an der University of Colorado in Boulder am 21. Oktober 1965.     

Membrane-bound F1 ATPase from Micrococcus Sp. ATCC 398E. Purificationa and characterization ny affinity chromatography.

A chemically reactive ATP analogue, 6-[(3-carboxy-4-nitrophenyl)thio]-9-beta-D-ribofuranosylpurine 5'-triphosphate (Nbs6ITP) has been synthesized. It has the ability to form stable thioether bonds between the 6-position of the purine ring and aliphatic mercapto groups. The nucleotide moiety of the reagent has been covalently bound to agarose, via iminobispropylamine and N-acetyl-homocysteine as space with the purpose of producing an affinity chromatography material. The affinity matrix binds solubilized F1 ATPase from a crude extract of Micrococcus sp. membranes. Afterwards the enzyme can be selectively eluted from the column at a defined ATP concentration. This method is superior to the conventional purification with respect to speed and convenince of the preparation. The affinity chromatography leads in a one-step process to the same purity to enzyme, substituting several steps of the conventional method. In addition, the affinity matrix was used for binding studies. Although the presence of Mg2+ ions is a prerequisite for the hydrolysis of nucleoside 5'-triphosphates, evidence is presented indicating that the binding of the nucleoside triphosphates to highly purified F1 ATPase from Micrococcus sp. appears not to be influenced by Mg2+ ion concentrations so far examined.