Decrease In ATP Content Research Articles

Effects of exercise and adrenaline on equine erythrocyte atp content

To investigate the claim that equine erythrocytes released from the spleen are older cells than those found at rest in the circulation, the 2,3, diphosphoglycerate (2,3 DPG), adenosine triphosphate (ATP) and creatine concentration of erythrocytes before and following splenic emptying were examined. Normal values for thoroughbreds (43) and ponies (10) at rest were established. Following either exercise or intravenous injection of adrenaline in six thoroughbreds, there was an increase in erythrocyte creatine content and a decrease in ATP concentration. Exercise produced a slight increase in 2,3 DPG while no change occurred with adrenaline. In two splenectomised ponies adrenaline only produced a decrease in erythrocyte ATP, with no change in creatine or 2,3 DPG content. It was concluded that erythrocytes released from the spleen are not aged cells. However, the stimuli used produced a decrease in ATP content which may affect the properties of the erythrocytes.

Modulation of mammalian sperm motility by quercetin

The flavonoid quercetin inhibits collective motility of ejaculated ram spermatozoa in the first 2 hr of incubation; during the next 3-4 hr motility is stimulated. To explain this interesting effect, we followed the influence of quercetin on sperm glycolysis, extracellular pH, ATP content, mitochondrial respiration, and lipid peroxidation. The collective motility of untreated cells is decreased to about 40% of the original motility during two hours of incubation. During this time, the rate of glycolysis is constant, respiration rate is increasing, there is no change in ATP content, the rate of lipid peroxidation is very slow, and the extracellular pH became very acidic (pH 5.5). It is concluded that motility is decreased due to this acidification. This acidification is prevented to some extent by quercetin, which indirectly inhibits glycolysis. Quercetin inhibits motility due to the inhibition of the plasma membrane calcium pump, as we showed previously (Breitbart et al., J Biol Chem 260:11548-11553, 1985). The motility of untreated cells is arrested after 3.5 hr of incubation, whereas quercetin-treated cells show high motility, which continues for additional 2-3 hr. After 3.5 hr, the control cells show no glycolytic activity, ATP content and respiration rates are decreased, and rate of lipid peroxidation is highly increased. At this time, quercetin-treated cells show no glycolytic activity, only a small decrease in ATP content and respiratory rate, and a very low rate of lipid peroxidation. Based on these data it is concluded that sperm motility after 3.5 hr of incubation is dependent mainly on mitochondrial respiration.(ABSTRACT TRUNCATED AT 250 WORDS)

The abnormal-shaped mitochondria in thymus lymphocytes treated with inhibitors of mitochondrial energetics

The effects of uncouplers (DNP, FCCP), oligomycin, and rotenone on the energetics and mitochondrial ultrastructure in lymphocytes have been studied. We confirmed the previous observations done on Ehrlich ascites and cardiomyocyte culture cells that uncouplers and respiratory inhibitors cause the appearance of ringlike and dumbbell-like mitochondria. It is shown that this effect does not correlate with decrease in ATP concentration, changes in oxygen consumption, or condensation of the mitochondrial matrix. FCCP (2 microM) is more effective in the induction of abnormal-form mitochondria than 240 microM DNP, oligomycin, or rotenone. Combined treatment with DNP, oligomycin, and rotenone or with DNP and rotenone produces an effect as strong as 2 microns FCCP. DNP (240 microM) and FCCP (2 microM) have a similar effect on respiration and intracellular ATP, but only the latter induces condensation of the mitochondrial matrix.

漁獲物の生きの保持に関する研究 ＩＩＩ 二，三海産魚における死後硬直の進行と貯蔵温度の影響

Yellowtail, bartailed flathead and Japanese striped knifejaw were spiked at the brain, and stored at O°and 10°C. Yellowtail set in rigor-mortis after 2h, irrespective of storage temperature, attaining the full-rigor state after 7h at O°C and 15h at 10°C. Rigor-mortis of bartailed flathead started after 10h at O°C and 13h at 10°C. This species reached the full-rigor state after 20h at O°C and 40h at 10°C. In the case of Japanese striped knifejaw, rigor-mortis began to appear immediately after killing at O°C whereas after 4h at 10°C. The full-rigor state was attained with this fish after 13 and 20h, respectively, in the above order of storage temperature. Therefore, rigor-mortis proceeded roughly two times slower at 10°C than at O°C, regardless of fish species. In accordance with rigor-mortis progress, the muscle of each species showed a 2-3 times slower decrease of ATP concentration when stored at 10°C than when stored at O°C. Disappearance of creatine phosphate preceded that of ATP while lactate accumulation was nearly maximal when ATP disappeared, irrespective of fish species and storage temperature. No marked differences in initial ATP concentration were observed among the fish species examined. The highest lactate level was encountered in yellowtail which exhibited the fastest progress of rigor-mortis at either storage temperature. Japanese striped knifejaw showed a much lower accumulation of lactate than yellowtail, the accumulation of which was even lower than that in bartailed flathead. On the other hand, rigormortis progress of Japanese striped knifejaw was comparable to that of yellowtail and somewhat faster than that of bartailed flathead.

Regulation of macrophage physiology by L-arginine: role of the oxidative L-arginine deiminase pathway.

The L-arginine content of the extracellular fluid in sites of predominant macrophage infiltration is reduced below plasma levels due to the activity of macrophage-derived arginase. Investigation of the effects of altered L-arginine availability on macrophage physiology reveals that culture of rat peritoneal macrophages in media containing L-arginine in the concentrations present in inflammatory lesions (less than 0.1 mM) enhances activation-associated functions. In contrast, culture in the higher L-arginine concentrations found in standard tissue culture media (0.4 to 1.2 mM) suppresses most macrophage functions (superoxide production, phagocytosis, and protein synthesis). An exception is the tumor cytotoxicity of Corynebacterium parvum-elicited macrophages which is enhanced by culture in supraphysiologic concentrations of L-arginine. Work reported here investigated the mechanisms for these L-arginine-dependent effects and, more specifically, the role of the recently described oxidative L-arginine deiminase pathway in the regulation of macrophage physiology. Overnight culture of resident or C. parvum-elicited peritoneal macrophages in media containing increasing concentrations of L-arginine (6 microM to 1 mM) resulted in: inhibition of electron transport chain activity (resident and C. parvum-elicited macrophages), increased lactate production (resident macrophages), and decreased ATP content (resident and C. parvum-elicited macrophages). In line with these findings, viability was markedly decreased after 2 days of culture when the initial L-arginine concentration was greater than or equal to 0.1 mM. As shown before, increasing media concentrations of L-arginine were associated with suppression of superoxide production and cytotoxicity in resident macrophages, and with reduced superoxide production and increased cytotoxicity in C. parvum-elicited macrophages. All L-arginine-dependent metabolic and functional alterations, as well as the loss of viability, were prevented by NG-monomethyl-L-arginine, a specific inhibitor of the oxidative L-arginine deiminase pathway. These results demonstrate that flux of L-arginine through the oxidative L-arginine deiminase pathway results in the inhibition of oxidative metabolism in rat macrophages. This metabolic inhibition may, through alterations in the macrophage high energy phosphate stores, mediate the suppression of cell functions and result ultimately in cell death.

Characterization of Scat (suture cataract), a dominant cataract mutation in mice

The autosomal, dominant mutation Scat (suture cataract) was found in ( ( 101 El × C3H El )F 1-hybrid mice. The severity of the cataract is dependent on the gene dose. The mutation causes an anterior suture opacity in heterozygotes amd microphthalmia with vacuolated lenses in homozygotes. In histological sections of lenses the heterozygotes exhibit a hydropic swelling of lens epithelium, whereas in homozygotes interruption and degeneration of lens fibers as well as clefts and folds of the capsule were observed. The mutation has a complete penetrance and constant expressivity. The body weight of the mutants is not altered; the mutation has no effects on fertility or viability. The lens wet and dry weights are diminished (more pronounced in the homozygotes). The water content of the lens is enhanced only in the homozygous Scat mutants. Biochemically, the lenticular content of water-soluble proteins is decreased in the homozygous Scat mutants. By electrophoresis, in the lenses of homozygous Scat mutants a different pattern of water-soluble proteins could be observed. The lenses of both, heterozygous and homozygous Scat mutants exhibit enhanced Na +,K +-ATPase activity and a decreased ATP concentration. The genetical, morphological or biochemical data suggest that the effect of the Scat mutation is distinct from other described cataract mutations in mice.

Nephrotoxicity assessment by measuring cellular ATP content: I. Substrate specificities in the maintenance of ATP content in isolated rat nephron segments

To clarify the characteristics of cellular ATP synthesis in individual nephron segments for assessing nephrotoxicity of chemicals, cellular ATP content was measured by the luciferin/luciferase system under various conditions using intact nephron segments isolated from male Sprague-Dawley rats. Increasing the duration of collagenase treatment of kidney slices significantly lowered the cellular level of ATP newly synthesized from 2 m m glutamine in PST at 37°C over 30 min ( p < 0.01). The tubular incubation time significantly affected the cellular ATP content in the early and middle portions (S 2) of the proximal tubule ( p < 0.05 and p < 0.01, respectively) over 20 min and in the late proximal tubule over 10 min. Among numerous substrates tested, such as d-glucose, glutamine, pyruvate, dl-lactate, and β-hydroxybutyrate, the substrate utilization for maintaining cellular ATP content was entirely variable according to each nephron segment. Pyruvate and glutamine were the best substrates in the proximal tubule. On the other hand, ATP production from glutamine was less than that from the other substrates in the distally located nephron segments: medullary and cortical thick ascending limbs of Henle's loop (MAL and CAL, respectively), distal tubule, cortical and medullary collecting tubules (CCT and MCT, respectively). In general, glucose, pyruvate, and lactate appear to be equivalent in maintaining ATP content in the distal segments of renal tubules. A monovalent cation ionophore, monensin, at 10 μg/ml decreased the cellular ATP content in MAL, CAL, and MCT significantly. Mercuric chloride (HgCl 2) was used as a model compound to study nephrotoxicity by investigating its effects on cellular ATP metabolism in microdissected nephron segments. HgCl 2 at 1 × 10 −6 m significantly decreased ATP content only in S 2 ( p < 0.05), clearly demonstrating S 2 to be the most sensitive segment within the nephron. These results indicate that measurement of cellular ATP content would be a useful method for forecasting the intrarenal toxic site and potency of possible nephrotoxic chemical compounds.

Effects of pH and ATP Concentrations on the Heat Contracture of Glycerinated Single Fibers of Rabbit Psoas Muscle

ABSTRACTThe occurrence of heat contracture of glycerinated single fiber of rabbit psoas muscle was investigated in relaxing solutions with varying pH values and ATP concentrations. The threshold temperature for inducing heat contracture of single muscle fibers in relaxing solutions decreases with increasing pH values and decreasing ATP concentrations. There seems to be a relationship between heat contracture of muscle fibers and the tremor of skeletal muscle of porcine stress syndrome. Also, there seems to be a relationship between the heat contracture and alkaline rigor development.

Biochemical and developmental features of experimental phenylketonuria induced by L-ethionine in suckling rats.

Suckling rats were injected subcutaneously with doses of L-ethionine (0.1 mumole/g body wt) at intervals of 12 hr. In the latter group, phenylalanine hydroxylase was effectively inhibited in vivo resulting in hyperphenylalaninemia and phenylketonuria. Due to the well-known sex-specific differences in L-ethionine metabolism female rats were much more affected by chronic administration of L-ethionine. The underlying mechanism of enzyme inhibition by ethionine could be disturbed protein synthesis and impaired protein phosphorylation, which was suggested by pronounced decreases in ATP content in liver. In the high dosage group depletions mainly of the branched-chain amino acids and lysine occurred in serum and brain, whereas the concentrations of methionine and tryptophan were increased. Tyrosine tended to be decreased in the course of hyperphenylalaninemia. Hyperphenylalaninemia and other resulting amino acid imbalances obviously impaired brain development during the early postnatal period. Concomitantly with reductions in protein concentrations, the activity of cathepsin D, a major intralysosomal acid proteinase, was increased in brain, suggesting also higher protein catabolism in brain. Side effects of this treatment, however, were higher mortality, loss of body weight, and a general impression of delayed development, resembling a state of undernutrition to some extent. These obvious side effects of ethionine limit the usefulness of ethionine as a suitable model for classic phenylketonuria in suckling rats.

Effects of idebenone on neurological deficits, local cerebral blood flow, and energy metabolism in rats with experimental cerebral ischemia

Improvement of energy metabolism in ischemic cerebral tissue benefits the therapy of occlusive cerebrovascular disorders. In the present study, the effects of 6-(10-hydroxydecyl)-2,3-dimethoxy-5-methyl-1, 4-benzoquinone (idebenone) on neurological signs, such as ischemic seizures, lactate and ATP contents of the cerebral cortex, and local cerebral blood flow, were assessed in stroke-prone spontaneously hypertensive rats (SHRSP) with experimentally induced cerebral ischemia. Experimental cerebral ischemia was caused by bilateral carotid artery occlusion (BCAO) in male SHRSP (8–10 weeks old). Pretreatment with idebenone (10–100 mg/kg, p.o.) for 3 or 10 days delayed the onset of ischemic seizure (acute stroke) and prolonged survival time in SHRSP roughly in a dose-dependent manner. When the compound (100 mg/kg, i.p.) was given once 30 min after BCAO, it exerted similar ameliorating effects on the neurological deficits. When idebenone (100 mg/kg for 3 days) was given orally, it did not significantly inhibit the decrease in regional cerebral blood flow induced by BCAO. However, the same treatment markedly inhibited increases in the lactate content and lactate/pyruvate ratio and the decrease in ATP content of the cerebral cortex. The compound did not affect cerebral blood flow in normal rats. These results suggest that idebenone ameliorates the neurological deficits related to cerebral ischemia, and that this effect is mediated by improving cerebral energy metabolism.

The flavonoid tangeretin inhibits invasion of MO4 mouse cells into embryonic chick heart in vitro.

Tangeretin, a flavonoid from citrus plants, was found to inhibit the invasion of MO4 cells (Kirsten murine sarcoma virus transformed fetal mouse cells) into embryonic chick heart fragments in vitro. The flavonoid appeared to be chemically stable in tissue culture medium, and the anti-invasive effect was reversible on omission of the molecule from the medium. Unlike (+)-catechin, another anti-invasive flavonoid, tangeretin bound poorly to extracellular matrix. It did not alter fucosylated surface glycopeptides of MO4 cells. Tangeretin seemed not to act as a microtubule inhibitor, as immunocytochemistry revealed no disturbance of the cytoplasmic microtubule complex. However, at anti-invasive concentrations of tangeretin, cell proliferation and thymidine incorporation appeared to be inhibited. When cultured on an artificial substrate, treated MO4 cells were less elongated, covered a larger surface area and exhibited a slower directional migration than untreated cells. From the decrease in ATP content in MO4 cells after tangeretin treatment, we deduce that this flavonoid inhibits a number of intracellular processes, which leads to an inhibition of cell motility and hence of invasion.

The use of the creatine kinase reaction to determine free energy change of ATP hydrolysis in anoxic cardiomyocytes

In isolated cardiomyocytes from adult rat heart the free energy change of ATP hydrolysis (dG) was determined under conditions of substrate-free anoxia. Changes of free cytosolic ADP concentrations, needed for the calculation of dG, were determined by two indirect methods since a direct measurement is not feasible: (i) via the mass action ratio of the creatine kinase reaction (CK) assuming near equilibrium conditions, and (ii) via quantification of the net hydrolysis of ATP to ADP by a detailed balancing of possible contribution to Pi production. Both approaches gave virtually identical results, showing that in anoxia only 6% of the ATP hydrolysed are hydrolysed to ADP and 94% completely to adenosine and further degradation products. The convergence of both methods also indicates that in this model the CK reaction is indeed catalysed near its equilibrium. Therefore estimations of free ADP and dG using its mass action ratio are valid. In anoxic cardiomyocytes dG values fell from 57 kJ/mol in normoxia to 42 kJ/mol after 120 min anoxia, corresponding to a decrease of ATP contents from 24 to 4 nmol/mg protein.

Erythrocyte osmotic fragility and cation concentrations during experimentally induced bovine anaplasmosis

1. 1. The osmotic fragility, the concentrations of Na, K and Ca, the osmolality and the total ATPase activity of bovine erythrocytes from uninfected and Anaplasma marginale-infected bovines were studied in an attempt to correlate these parameters with the decrease in the cellular ATP concentration reported during bovine anaplasmosis. 2. 2. The osmotic fragility found in infected bovine erythrocytes, at 0.52% NaCl, was about two times greater than that observed in non-infected bovines. The increase in osmotic fragility was directly related to the increase in intra-erythrocytic parasitemia. 3. 3. The decrease in ATP concentration reported during bovine anaplasmosis could not be directly related to the increased fragility of these cells. The artificial depletion of erythrocytic ATP did not reproduce the same alteration in the osmotic response to NaCl. 4. 4. The plasmatic and cytoplasmatic concentrations of Na, K and Ca did not change significantly during bovine anaplasmosis, whereas the interior of the erythrocytes became hyperosmolal. 5. 5. A. marginale-infected bovine erythrocyte membranes showed an increased ATPase activity when compared to control bovines. Parasite-enriched fractions also presented ATPase activity.

Inhibition by calcium of beta adrenoceptor mediated cAMP responses in isolated bovine corneal epithelial cells

The increases in adenosine 3',5' monophosphate (cAMP) content were measured in isolated bovine corneal epithelial cells in response to either adrenergic agonists or adenylate cyclase stimulation. The beta selective adrenergic agonist, isoproterenol, and the adrenergic agonists, norepinephrine as well as epinephrine elicited large increases in cAMP accumulation. At their maximum effective concentrations, the respective increases were 16-fold, 6.6-fold and 4.7-fold. These stimulatory effects were completely inhibited by the beta selective adrenergic antagonist, propranolol. Similarly forskolin increased cAMP content more than 3-fold. These increases and the previous identification of beta adrenoceptors in fresh intact bovine corneas as well as cells in culture indicate that this enzymatic dissociation procedure does not affect the cAMP responses to either adrenergic agonists or forskolin. The relationship was considered between increases in Ca2+ concentration and the effects of either isoproterenol or forskolin, on cAMP accumulation. There were no changes in any of the cAMP responses at bathing solution Ca2+ concentrations between 0.01 microM and 1 microM. However, in cells permeabilized to Ca2+ with 10 microM ionomycin, increases within this concentration range depressed the baseline levels of cAMP content. Furthermore, the stimulatory effects of both forskolin and isoproterenol on cAMP accumulation were significantly blunted in this concentration range. These blunting effects by Ca2+ were not the result of any measurable decrease in ATP content. This negative relationship between increases in Ca2+ concentration and increases in cAMP content indicates that changes in intracellular Ca2+ concentration could modulate the second messenger function of cAMP linked to these agents.

Efficiency of work performance and contraction velocity in isotonic tetani of frog sartorius.

Mechanical work, ATP, ADP, PC, free creatine and lactate concentrations were determined on IAA poisoned frog sartorii tetanically stimulated in humidified N2 at 10 degrees C in isotonic conditions (0.25 or 0.45 Po). Tetanus duration was 0.35 s, number of tetani was varied from 0 (rest) to 25 (exhaustion). The mechanical work performed per mole ATP + PC split (W*P) amounted on the average to 16.7 kJ/mol. It was observed, however, that W*P increased from about 13 to about 24 kJ/mol with decreasing ATP concentration from about 2 (resting value) to about 1 mumol/g and that this decrease in ATP was associated with a decrease of the shortening (and relaxation) speed of the muscle to about 30% of the values observed on the first tetanus. It is concluded that the thermodynamic efficiency of muscle contraction, calculated from the ratio of WP* (measured) to the thermodynamic affinity (free energy change) of ATP hydrolysis (estimated) increases from about 0.3 to about 0.5 with decreasing ATP concentration and shortening speed.

Hepatic autophagy and intracellular ATP a morphometric study

In order to estimate the sensitivity of macroautophagy in liver toward changes in ATP we have analyzed the volume density of the autophagic/lysosomal system in isolated rat hepatocytes, incubated under conditions where intracellular ATP was partially depleted, (a) It appeared that reduction of the intracellular ATP concentration by 30–50% decreased the volume density of autophagic vacuoles by 70%. (b) Partial ATP depletion did not involve significant changes in the volume density of dense bodies. Together with studies showing that the rate of overall proteolysis via macroautophagy decreases with decreasing ATP concentration ( P. J. A. M. Plomp, E. J. Wolvetang, A.K. Groen, A. J. Meijer, P. B. Gordon, and P. O. Seglen (1987) Eur. J. Biochem. 164, 197–203) our data indicate that changes in intracellular ATP primarily affect early steps in the autophagic/proteolytic pathway.

The effect of inhibition of mitochondrial energy metabolism on the transient inward current of isolated guinea-pig ventricular myocytes

Delayed afterdepolarizations (DADs) might underlie ischemic or reperfusion arrhythmias (Ferrier et al., 1985; Coetzee and Opie, 1987). These DADs are the result of a transient inward current iti, which is caused by instability of the intracellular level of free Ca2+ (Cai) due to an oscillatory release of Ca from the sarcoplasmic reticulum (Kass et al., 1978). DADs are known to be abolished by hypoxia and by metabolic inhibition (Di Gennaro et al., 1987; Coetzee and Opie, 1987), which could be caused by a number of different mechanisms: (1) The large increase of potassium conductance associated with metabolic inhibition (Vleugels et al., 1980; Isenberg et al., 1983) could prevent iti from causing a marked depolarization, and would thus "mask" the DADS. (2) Although metabolic inhibition will eventually result in an increase of Cai, a temporary decrease could initially take place, thereby minimizing the Ca instability. Two mechanisms are known which might produce such an effect: Firstly, the shortening of the action potential which occurs during metabolic inhibition will markedly reduce the time during which Ca channels remain open, thereby causing a diminished total Ca influx during the action potential (Isenberg et al., 1983; Noma and Shibasaki, 1985; Kakei et al., 1985). Secondly, a direct reduction of iCa by a decrease in ATP concentration, described by Irisawa and Kokubun (1983), could also contribute to a decreased Ca load. (3) Metabolic inhibitors could possibly interfere with the cycling of Ca between different compartments within the cell, thereby altering the temporal variation in Cai, and thus influencing iti.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of Bethanechol on Glycolysis and High Energy Phosphate Metabolism of the Rabbit Urinary Bladder

The urinary bladder, similar to other smooth muscles, utilizes glucose as one of its primary sources of metabolic energy. We have studied the effect of bethanechol on both glycolysis and high energy phosphate metabolism. The results can be summarized as follows: bethanechol administration in vitro stimulates a 30% decrease in intracellular glycogen, a 100% increase in lactic acid production, and an 80% increase in CO2 generation. Although there was a rapid and sustained decrease in the intracellular concentration of creatine phosphate, there was only a minor decrease in the intracellular concentration of ATP. There were no changes in adenine uptake or de novo ATP synthesis.

Stepwise sliding disintegration of tetrahymena ciliary axonemes at higher concentrations of ATP

AbstractSeveral characteristics of the sliding disintegration of Tetrahymena ciliary axonemes were found by turbidimetric assay, the ATP‐regenerating system, and quantitative determination of the ATP concentration. At ATP concentrations exceeding 40 μM, the response in terms of turbidity was biphasic and could be divided into three phases. The dependence of each phase on ATP concentration was examined. The time duration of phase 2 increased with ATP concentration. When the ATP concentration was kept constant by the ATP‐regenerating system, consisting of pyruvate kinase and phosphoenol pyruvate, the time duration of phase 2 increased with the concentration of phosphoenol pyruvate. On examining the change in turbidity with decreasing ATP concentration, the transition from phase 2 to phase 3 was found to occur at an ATP concentration of 40 μM.Dark‐field and electron microscopy indicated the sliding disintegration to be closely correlated with the degree of tubidity. At phase 1, one or two doublets extruded from most of the axonemes, and disintegration failed to progress during phase 2. At the transition point from phase 2 to 3, at about 40 μM, ATP, other doublets were noted to extrude from the axonemes one after the other, causing turbidity to be minimal by the end of phase 3.The ATP concentration dependence of stepwise sliding disintegration suggests that each axoneme may possess the ability to regulate doublet microtubule sliding at lower or higher concentrations of ATP. In response to local differences or gradients of ATP concentration along the axoneme, the axonemes may cause localized sliding of doublets, thus subsequently generating active bending movement.

Characterization of the bacteriophage T3 DNA packaging reaction in vitro in a defined system

The bacteriophage T3 DNA packaging system in vitro defined here is composed of purified proheads and two non-capsid proteins, the products of genes 18 and 19 (gp18 and gp19). In this system, a precursor complex (50 S complex) accumulates in the presence of adenosine 5′- O-(3′-thiotriphosphate) (ATP-γ-S), a non-hydrolyzable analog of ATP. The 50 S complex is converted to a filled head in the presence of ATP. The conversion of the 50 S complex, formed by preincubation with ATP-γ-S, to the mature head proceeds in a synchronous manner after the addition of ATP. The lag time for formation of mature heads from the 50 S complex is 1.8, 4.5 and 6.8 minutes at 30, 25 and 20 °C, respectively. DNA is translocated into the capsid at a constant rate of 5.7 × 10 3 base-pairs per minute at 20 °C. The conversion of the 50 S complex to the mature head exhibits a sigmoidal relationship with respect to the concentration of ATP. the concentration for half-maximal activity being about 20μ m. The transition of the prohead to the expanded capsid occurs at 20 °C at one minute 40 seconds after the initiation of DNA translocation, when one-fourth of the genome has been packaged into a prohead. At the same time, the capsid-DNA complex becomes stable to high concentrations of salt. When DNA translocation is interrupted by the addition of ATP-γ-S, packaged DNA exits at 0 °C as well as at 20 °C but the exit of DNA stops after one-third of the genome is inside the capsid. After exit, DNA is retranslocated into the expanded capsid by the addition of ATP at a rate of about 5.7 × 10 3 base-pairs per minute at 20 °C. The decrease in concentration of ATP interrupts DNA translocation into the capsid but does not induce DNA exit. Interrupted DNA translocation may be reinitiated by the addition of ATP. DNA exit is not induced by the addition of ATP-γ-S to mature heads or partially filled heads pretreated with DNase.