Inhibition of E coli ATPase activity by a troponin component, TN-I, and by mitochondrial ATPase inhibitor.

The enzymic activity of Mg2+- or Ca2+-stimulated ATPase from Escherichia coli was inhibited by one of the troponin components, TN-I, and by mitochondrial ATPase inhibitor (F1-inhibitor). The inhibitory ability of component TN-I against Mg2+-stimulated AtPase activity was lost after digestion of component TN-I with trypsin. The Mg2+-stimulated ATPase activity inhibited by component TN-I was completely restored by the addition of another troponin component TN-C.     

Inhibition ofE. coli ATPase activity by a troponin component,TN-I,and by mitochondrial ATPase inhibitor

Summary The enzymic activity of Mg²⁺-or Ca²⁺-stimulated ATPase fromEscherichia coli was inhibited by one of the troponin components, TN-I, and by mitochondrial ATPase inhibitor (F₁-inhibitor). The inhibitory ability of component TN-I against Mg²⁺-stimulated ATPase activity was lost after digestion of component TN-I with trypsin. The Mg²⁺-stimulated ATPase activity inhibited by component TN-I was completely restored by the addition of another troponin component, TN-C.     

Na, K ATPase activity during early postnatal development of the rat submandibular gland

The activity of the ouabain-sensitive Na, K ATPase was measured in membrane fractions of the submandibular gland of 1-, 7-, 14- and 21-day-old rats. This activity increased with age and reached adult levels by 21 days.     

Na,K ATPase activity during early postnatal development of the rat submandibular gland

Summary The activity of the ouabain-sensitive Na, K ATPase was measured in membrane fractions of the submandibular gland of 1-, 7-, 14- and 21-day-old rats. This activity increased with age and reached adult levels by 21 days.     

Effect of cholesterol oxidation on (Na+, K+) ATPase activity of erythrocyte membranes.

By incubation of human erythrocyte ghosts with cholesterol oxidase (EC 1.1.3.6) part of the cholesterol of the membrane is replaced by 4-cholesten-3-one. This alteration in the sterol composition is accompanied by an inhibition of the (Na+, K+) ATPase of the erythrocyte membrane.     

The plasma membrane proton-translocating ATPase

Living cells require membranes and membrane transporters for the maintenance of life. After decades of biochemical scrutiny, the structures and molecular mechanisms by which membrane transporters catalyze transmembrane solute movements are beginning to be understood. The plasma membrane proton-translocating adenosine triphosphatase (ATPase) is an archetype of the P-type ATPase family of membrane transporters, which are important in a wide variety of cellular processes. The H+-ATPase has been crystallized and its structure determined to a resolution of 8 angstrom in the membrane plane. When considered together with the large body of biochemical information that has been accumulated for this transporter, and for enzymes in general, this new structural information is providing tantalizing insights regarding the molecular mechanism of active ion transport catalyzed by this enzyme.     

Effect of cholesterol oxidation on (Na+, K+) ATPase activity of erythrocyte membranes

Summary By incubation of human erythrocyte ghosts with cholesterol oxidase (EC 1.1.3.6) part of the cholesterol of the membrane is replaced by 4-cholesten-3-one. This alteration in the sterol composition is accompanied by an inhibition of the (Na⁺, K⁺) ATPase of the erythrocyte membrane.Acknowledgments. This study was supported by the Deutsche Forschungsgemeinschaft Bonn-Bad Godesberg and the Sonderforschungsbereich 90 of the University of Heidelberg. We thank Mr.J. Schmidt and MissI. Geldmacher for skilful technical assistance.     

ATPase activity in mercury intoxicated eels

Summary Eels intoxicated by lethal doses of HgCl₂ accumulate mercury in their gills. Mercury inhibits the ouabainsensitive Na+K+ATPase activity of gills involving the rupture of the fish NaCl balance.The author wishes to express his sincere gratitude to Professor A. Disteche and Professor E. Schoffeniels for discussions during the investigation. This research was carried out in participation in the Belgian National Research and Development Program on the Environment-Water-Sea Project-Office of the Prime Minister-Interministerial Commission for Science Policy and Programmation.     

Age dependent changes in Na+−K+, activated ATPase activity of locust rectum

Summary Na⁺–K⁺, activated ATPase showed an increase in activity at the beginning of the 5th instar and adult life. This was followed by a relatively constant level of activity which, in larval preparations only, was not maintained but decreased with the onset of metamorphosis.     

(Ca−Mg) ATPase activity of human erythrocyte membranes: Influence of incubation buffer

Summary Activity of the (Ca–Mg) ATPase of human red blood cell membranes is highly dependent on the specific buffer used in the ATPase assay. Activity is highest in histidine and/or imidazole buffers and is lowest in HEPES buffer.This research was supported by Public Health Service Grants AH-16436 and GM 00109-16. We wish to acknowledge the useful suggestions of Thomas R. Hinds and the excellent clerical assistance of Ms Laurie Asher.     

Membrane fractions of rabbit granulocytes. 1. The presence of ouabain-sensitive ATPase and the electrophoretic patterns.

In one or two membrane fractions isolated from the homogenate of casein-induced rabbit granulocytes ouabain-sensitive ATPase activity was found definitely. Electrophoretic patterns of these membrane fractions showed no significant differences in staining for carbohydrate and protein.     

Induction of chromosomal aberrations by the anthracycline antitumor antibiotics N,N-dimethyldaunomycin and aclacinomycin A

Summary The clastogenic effect of the anthracycline antitumor antibiotics, N,N-dimethyldaunomycin and aclacinomycin A, was studied in a murine hemopoietic cell line (Friend leukemia cells). A dose-dependent increase in chromatid lesions, i.e., achromatic lesions, chromatid breaks, chromatid deletions and triradial or quadriradial chromosomal exchange figures, was found. It appears that the clastogenicity of N,N-dimethyldaunomycin and aclacinomycin A is lower than that of the classic anthracycline, daunomycin, which is also a potent mutagen and carcinogen. The data demonstrate that the capacity of chemicals to induce point mutations and chromosomal aberrations may not necessarily be correlated: aclacinomycin A is devoid of mutagenic activity in bacterial (Salmonella typh.) and mammalian cell (HGPRT) mutagenesis assays, and is non-carcinogenic in rats. Nevertheless, it was now found to possess clastogenic activity.     

The activation of membrane ATPase by ouabain in several tissues of the golden hamster (Mesocricetus auratus) in the absence of potassium

Summary The activity of the membrane ATPase of 5 organs of the golden hamster was increased by 10^–7–10^–3 moles/l ouabain in K⁺-free medium. In similar experiments on rats no increase was observed.     

Purification of a membrane protein modifying the sensitivity of Na+/K+ ATPase to ouabain from a supernatant of plasma membrane treated with EDTA]

A membrane protein of M. W. 32,000 D and pI 5 has been purified from EDTA-treated plasma membrane supernatants by Page without detergents. These membranes are purified from the MF2S cell line issued from the murine plasmacytoma MOPC 173. The purified protein was able to induce a 300 fold increase in the Na+/K+ ATPase resistance to ouaba?n thus mimicking the original resistance of the enzyme to the drug.     

Skinned smooth muscle: Time course of force and ATPase activity during contraction cycle

Summary The time course of ATPase activity and force has been determined during contraction and relaxation in skinned (hyperpermeable) anterior byssus retractor muscle, ABRM, ofMytilus edulis and compared with corresponding measurements on skinned taenia coli of guinea-pigs. Following a calcium-induced contraction, lowering the [Ca⁺⁺] to 10^–8 M rapidly reduces ATPase activity within 2 min to resting levels while force declines only to about 30–50% of maximal tension within the same time. Thus slow relaxation is due to a catch-like-state which is common to different kinds of smooth muscles and can be reduced with cAMP in ABRM and by P_i in taenia coli.The support by the Deutsche Forschungsgemeinschaft and the excellent technical assistance of Mrs K. Winnikes are gratefully acknowledged.     

The response to potassium of the Na-K pump ATPase in low-K red blood cells from cattle at birth and in later life

It is shown that in low-K red blood cells of cattle the apparent affinity for K(1/Kapp K) at an inhibitory site of the Na-K ATPase increases markedly during the first 3 months of life. This site probably is the Na accepting site at the internal membrane surface and the change in Kapp K reflects an increase in KNa/KK, the ratio of the true dissociation constants. This effect may explain the concomitant fall in cellular K concentration.     

Induction of chromosomal aberrations by the anthracycline antitumor antibiotics N,N-dimethyldaunomycin and aclacinomycin A

The clastogenic effect of the anthracycline antitumor antibiotics, N,N-dimethyldaunomycin and aclarcinomycin A, was studied in a murine hemopoietic cell line (Friend leukemia cells). A dose-dependent increase in chromatid lesions, i.e., achromatic lesions, chromatid breaks, chromatid deletions and triradial or quandriradial chromosomal exchange fiqures, was found. It appears that the clastogenicity of N,N-dimethyldaunomycin and aclacinomycin A is lower than that of the classic anthracycline, daunomycin, which is also a potent mutagen and carcinogen. The data demonstrate that the capacity of chemicals to induce point mutations and chromosomal aberrations may not necessarily be correlated: aclacinomycin A is devoid of mutagenic activity in bacterial (Salmonella typh.) and mammalian cell (HGPRT) mutagenesis assays, and is non-carcinogenic in rats. Nevertheless, it was now found to possess clastogenic activity.     

Norepinephrine-sensitive Na+/K+ ATPase activity in brown adipose tissue

Zusammenfassung Norepinephrin fördert in vitro die Tätigkeit einer Ouabain empfindlichen Na⁺/K⁺ ATPase. Diese Wirkung, durch eine geringe Dosis eines-adrenergischen Antagonisten eingeschränkt, scheint offenbar von einer 3,5-zyklischen AMP unabhängig zu sein. Die Tätigkeit der Brenzkatechin-induzierten ATPase stimmt mit der Auffassung überein, dass im aktivierten braunen Fettgewebe der Energieumsatz zusammen mit dem vermehrten Sauerstoffverbrauch ansteigt.

---

---

Supported in part by research grants NASA No. NGR-05-004-035 and USPHS No. HD-03268. P.A.H. is a PHS predoctoral fellow (No. 1-GM-33, 505-04). A preliminary report of this work was presented at the 1969 Fall meeting of the Am. Physiol. Society; an abstract of this talk is contained in The Physiologist12, 257 (1969). We thank J. F.Detrick for his technical assistance.     

Effect of heat treatment on the ATPase activity of various sarcoplasmic reticulum preparations

Summary Ca²⁺-stimulated ATPase activity of sarcoplasmic reticulum (SR) preparations is activated after a short period of preincubation at temperatures between 40 and 45°C, but for temperatures higher than 48°C pronounced denaturation is observed. Heat denaturation is decreased if Mg²⁺ or K⁺ are present during heat treatment.Acknowledgments. This work was supported by research grants from Instituto de Alta Cultura (Grant No. CB/2) and the Calouste Gulbenkian Foundation. We are grateful to Drs.A. P. Carvalho andV. M. C. Madeira for many discussions and suggestions during the course of this work.     

Effect of heat treatment on the ATPase activity of various sarcoplasmic reticulum preparations.

Ca2+-stimulated ATPase activity of sarcoplasmic reticulum (SR) preparations is activated after a short period of preincubation at temperatures between 40 and 45 degrees C, but for temperatures higher than 48 degrees C pronounced denaturation is observed. Heat denaturation is decreased if Mg2+ or K+ are present during heat treatment.