The role of protein surface charges in ion binding

Protein engineering is a means of probing the role of electrostatic interactions in protein functions; this elegant technique has been applied to the elucidation of electrostatic effects in enzyme catalysis. Here we show how the use of mutant proteins allows the determination of the contributions of individual charges to the free energy of ion binding to proteins. We have investigated the importance of three negatively charged side chains in the binding of Ca2+ to bovine calbindin D9K (ref.2): these are clustered around the calcium sites but are not directly involved as ligands. Each of these charges is found to contribute approximately 7 kJ mol-1 to the free energy of binding of two Ca2+ ions and to affect the cooperativity of Ca2+ binding. The influence of surface charges on ion binding to proteins may be more common than generally supposed and could have important consequences for protein function.     

The angiotensin converting enzyme inhibitor enalapril in acute ischemic renal failure in rats

The influence of the renin-angiotensin system on renal hemodynamics, tubular pressure and tubulo-glomerular feedback was investigated with the angiotensin converting enzyme inhibitor MK 421 (enalapril), in uninephrectomized rats with and without ischemia-induced acute renal failure. In animals with normal renal function proximal tubular pressure and tubulo-glomerular feedback response were lowered by enalapril long-term treatment, whereas glomerular filtration rate and renal blood flow were not influenced by the drug. After 45 and 70 minutes ischemia there was no difference between treated and untreated animals in the severely impaired glomerular filtration rate. Renal blood flow remained unaffected by the treatment. The histological damage due to ischemia (tubular casts, tubular necrosis and medullary capillary congestion) was not influenced by enalapril. As tubulo-glomerular feedback had been significantly inhibited during renin-angiotensin inhibition, its importance in mediating acute renal failure remains doubtful; other factors such as tubular obstruction and medullary congestion may be crucial.     

Construction and use of human chromosome jumping libraries from NotI-digested DNA

A basic difficulty in the molecular analysis of genes identified by mutations in the mammalian genome is the need to cover genetic distances corresponding to several hundred kilobases or more by molecular techniques like chromosome walking. In chromosome jumping, this limitation is overcome by the deletion of all but the extreme ends of large DNA molecules before cloning. We describe here the construction and characterization of a NotI 'jumping library' from human DNA. To characterize this library, random clones were analysed by restriction mapping. Clones carrying unique end fragments were characterized further by hybridization to Southern blots of NotI-cleaved human DNA separated on pulsed field gradient (PFG) gels. As a first step in a directional walk, the library was screened with a clone containing a NotI site cleaved in genomic DNA ('NotI linking clone') localized to the distal third of the short arm of human chromosome 4 (A.-M.F. & T.P., unpublished data). Starting and end points of two identified clones were positioned within a restriction map covering 850 kilobases.     

Vitamin K-dependent blood coagulation proteins form hetero-dimers

The later stages of the blood coagulation cascade are characterized by the presence of vitamin K-dependent proteins and their involvement in membrane-bound, multi-protein converting complexes with an essential requirement for calcium ions. Specific interactions between zymogens and activating enzymes have not yet been identified. Here we describe a crystallographic study of prothrombin fragment 1 (residues 1-156 of prothrombin) which indicates that vitamin K-dependent coagulation proteins have specific association sites that allow them to form hetero-dimers. The calcium-induced formation of a hetero-dimer between fragment 1 and factor X is demonstrated by cross-linking. Such hetero-dimers of vitamin K-dependent proteins could be significant in the coagulation system.     

Some interesting points on the histochemistry of the cytoplasmic inclusions of the ageing spinal neurones of reptiles,birds and mammals

Résumé L'analyse histochimique comparée des neurones des Reptiles, des Oiseaux et des Mammifères révèle que les divers corps qui y sont inclus ont une histochimie identique. Les sphéroïdes lipides abondent dans les phospholipides des jeunes neurones qui sont transformés en corps triglycérides chez les adultes. Les corps lipides ne sont pas un produit de sécrétion de l'appareil canaliculaire de Golgi dont l'homologue a été observé seulement dans les neurones adultes. La substance de Nissl est pleine de RNA.     

Beobachtung eines reversiblen Komproportionierungseffektes unter Lichteinwirkung bei Chrom-tris-2,2′-dipyridyl-Komplexen

Summary The effect of light and dark upon comproportionation and disproportionation respectively of chromium tris-bipyridine complexes is described.     

Local metabolic response to physio-pathological demands: The pentose phosphate pathway

Résumé On a constaté dans le métabolisme de glucose une augmentation de l'utilisation de la voie oxydative directe dans la paroi de l'estomac durant la digestion, et dans celle des vaisseaux sanguins pendant inflammation, réparation et reconstruction. Les resultats ont été analysés en fonction de leur intérêt pour les problèmes cliniques.

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Investigation was conducted during tenure of Special Research Fellowship (No. 11091), U.S. Public Health Service.     

Alkaloide ausSchizozygia caffaeoides (Boj.) Baill.

Summary FromSchizozygia caffaeoides (Boj.) Baill. (Apocynaceae), one major and ten minor new alkaloids have been isolated and characterised. Most of them are N-acylindoline derivatives, partly with the hitherto unknown 5,6-methylendioxyindoline system. Some possible correlations between these alkaloids are discussed.

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Den Herren Dres.H. Fritz, R. W. Schmid undH. Wagner und ihren Mitarbeitern danken wir für Spektren und Mikroanalysen.     

Thyroid-catecholamines interrelationships in free fatty acids release from adipose tissue

Zusammenfassung Die Abgabe freier Fettsäuren aus dem mesenterialen Fettgewebe kann durch Phentolamin nicht nur bei hyperthyroeten, sondern auch bei euthyreoten Ratten blockiert werden. Die erhöhte Freisetzung bei Hyperthyreose lässt sich wahrscheinlich durch eine gesteigerte Katecholaminaktivität erklären.     

Sex chromatin as a marker in some rabbit cells

Zusammenfassung In frisch isolierten Kaninchenfibroblasten-und Leukocyten kann kein Geschlechtsdimorphismus festgestellt werden. Nach einigen Tagen Zellkultivierung findet man in 84–93% (durchschnittlich 91,5%) der weiblichen Fibroblasten den Geschlechtschromatinkörper. Nur 0–2% (durchschnittlich 0,5%) der männlichen Fibroblasten weisen ein dem Geschlechtschromatinkörper ähnliches Gebilde auf. In Leukocytenkulturen sind die entsprechenden Zahlen 82–90% (durchschnittlich 89%) für weibliche, 0–1% (durchschnittlich 0,3%) für männliche Zellen. Diese Daten lassen den Geschlechtschromatinkörper als brauchbaren Zellmarkierer beim Kaninchen erscheinen.