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PubMed1985

Isolation and characterization of four forms of trehalase from rabbit kidney cortex.

Four forms of renal trehalase were isolated and purified to homogeneity. Hydrophobic interaction chromatography separated two forms; A-form and B-form. Both forms were subdivided further on Con A-Sepharose and were stained with periodic acid-Schiff reagent, indicating that they are glycoproteins. The four forms of renal trehalase showed no significant difference in Km values for trehalose and K1 values for various inhibitors. The optimum pH of the four forms was pH 6.0 in phosphate buffer. Apparent molecular weights on gel filtration of the four forms were the same, 175,000. Furthermore, the four forms showed the same antigenicity on double immunodiffusion. However, isoelectric point (pI), susceptibility to HgCl2, stability at -80 degrees C and Na+ activation behavior were different. Glycoprotein forms were more susceptible to HgCl2 and showed lower Na+ activation than nonglycoprotein forms. The pI of less hydrophobic forms (A1, A2) was more acidic than that of more hydrophobic forms (B1, B2). On the basis of these results, it is likely that four forms of renal trehalase are "isozymes."

Animals↗

PubMed1976

Isolation of renin granules from rat kidney cortex and evidence for an inactive form of renin (prorenin) in granules and plasma.

Renin was demonstrated in particles having a sedimentation velocity similar to that of mitochondria during differential centrifugation separated renin granules from the bulk of mitochondria and lyosomes, as well as from microsomes and cytoplasm. The density of renin granules was 1.202, which differed from the mean equilibrium densities of mitochondria (1.175) and lysosomes (1.170 and 1.230) in the heavy granule fraction. In studies involving gel filtration and polyacrylamide gel electrophoreis, renin granules appeared to contain an inactive form of renin that could be activated by acid treatment, had a higher apparent molecular weight than renin, and may be a more basic molecule. Inactive renin was also studied in plasma by electrophoresis and may originate from renin granules after exocytosis by the juxtaglomerular cells. Inactive renin may be a biosynthetic precursor (prorenin) and may be activated within the cell by a specific protease consequent upon the fusion of renin granules with lysosomes, thus providing a mechanism for the rapid regulation of renin activity prior to secretion.

Animals↗

PubMed1975

Distribution of parathyroid hormone-stimulated adenylate cyclase in plasma membranes of cells of the kidney cortex.

Free flow electrophoresis was employed to separate renal cortical plasma membranes into luminal (brush border microvilli) and contraluminal (basal-lateral membrane) fractions. During the separation adenylate cyclase activity was found to parallel the activity of Na+-K+-activated ATPase, an enzyme which is present in contraluminal but not in luminal membranes. In the basal-lateral membrane fraction the specific activities of adenylate cyclase and Na+-K+-activated ATPase were 4.4 and 4.6 times greater, respectively, than in the brush border fraction. The adenylate cyclase of the basal-lateral membrane fraction was specifically stimulated by parathyroid hormone which maximally increased enzyme activity eightfold. The biologically active (1-34) peptide fragment of paratyhroid hormone produced a 350% increase in adenylate cyclase activity. In contrast, calcitonin, epinephrine and vasopressin maximally stimulated the enzyme by only 55, 35 and 30%, respectively. These results indicate that adenylate cyclase, specifically stimulated by parathyroid hormone, is distributed preferentially in the contraluminal region of the plasma membrane of renal cortical epithelial cells.

3',5'-Cyclic-AMP Phosphodiesterases↗

PubMed1983

Isolation of basolateral and brush-border membranes from the rabbit kidney cortex. Vesicle integrity and membrane sidedness of the basolateral fraction.

A rapid and reproducible method has been developed for the simultaneous isolation of basolateral and brush-border membranes from the rabbit renal cortex. The basolateral membrane preparation was enriched 25-fold in (Na+ + K+)-ATPase and the brush-border membrane fraction was enriched 12-fold in alkaline phosphatase, whereas the amount of cross-contamination was low. Contamination of these preparations by mitochondria and lysosomes was minimal as indicated by the low specific activities of enzyme markers, i.e., succinate dehydrogenase and acid phosphatase. The basolateral fraction consisted of 35-50% sealed vesicles, as demonstrated by detergent (sodium dodecyl sulfate) activation of (Na+ + K+)-ATPase activity and [3H]ouabain binding. The sidedness of the basolateral membranes was estimated from the latency of ouabain-sensitive (Na+ + K+)-ATPase activity assayed in the presence of gramicidin, which renders the vesicles permeable to Na+ and K+. These studies suggest that nearly 90% of the vesicles are in a right-side-out orientation.

Adenosine Triphosphatases↗

PubMed1983

Carrier-mediated transport of amino-cephalosporins by brush border membrane vesicles isolated from rat kidney cortex.

The uptake of cephalosporin antibiotics by brush border membrane vesicles isolated from rat renal cortex has been studied by a rapid filtration technique, demonstrating a carrier-mediated transport system for amino-cephalosporins such as cephalexin and cephradine. The antibiotics were taken up into an osmotically reactive intravesicular space. The uptake of cephalexin was saturable (apparent Km2.2 mM), was inhibited by structural analogues and sulfhydryl reagents, and was stimulated by the countertransport effect, although the Na+ gradient did not affect the uptake. This transport system was essentially different from the transport system for p-aminohippurate in brush border membranes. The uptake properties for cephradine in brush border membrane vesicles appeared to be similar to those for cephalexin. The present results suggest the existence of a carrier-mediated transport system for amino-cephalosporins in brush border membranes. This system may be a part of the mechanism of tubular reabsorption of these antibiotics.

Animals↗

PubMed1986

Ca2+-dependent ATPases in the basolateral membrane of rat kidney cortex.

The basolateral segment of the rat renal tubular plasma membrane possesses Ca2+-dependent ATPase activity which was independent of Mg2+. Two kinetic forms were found: one, was a high affinity (apparent Km for free Ca2+ of 172 nM) low capacity (Vmax of 144 nmol of Pi X min-1 mg-1 protein) type; the other, had low affinity (apparent Km of 25 microM) and high capacity (896 nmol of Pi X min-1 X mg-1 protein). Mg2+ inhibited both Ca2+-ATPases. The high affinity enzyme exhibited positive cooperativity with respect to ATP, with a n value of 1.6. Ca2+-ATPase activity was not affected by calmodulin and was not inhibited by vanadate. On the other hand, both high and low affinity Ca2+-ATPase activities were increased when 1,25-dihydroxycholecalciferol was given to vitamin D-deficient rats. Kinetically, the enhanced activities were due to an increase in the Vmax values; the apparent affinities for free Ca2+ were not changed. The physiological function of the vitamin D-sensitive, Mg+-independent, Ca2+-ATPase activities remains to be established.

Adenosine Triphosphate↗

PubMed1981

Effect of hypo- and hypertonic saline and raffinose on cell content of Na, Cl and K in respiring rat kidney cortex at 37 degree.

Changes in cell Na, K and Cl content (per kg dry weight) and concentration (per kg intracellular water) were measured in respiring rat renal cortical slices incubated for 60 min at 37 degree in hypo- and hypertonic saline with and without the addition of a non-electrolyte (raffinose). Both hypertonic saline and raffinose increased cellular concentration of Na, Cl and K, the former by producing minimal cell shrinkage and major entry of Na and Cl into the cell, the latter entirely by cell shrinkage leading also to a loss of Na content but not of K. In hypotonic saline both content and concentration of Na and Cl did not change significantly, whereas that for K dropped markedly. Addition of ouabain (1 mmol/1) produced a significant gain of cell Na and loss of K on a one to one basis but did not alter the effect of raffinose. It is concluded that sustained changes in cell ion concentrations and content in response to pericellular osmolality are produced and that these are directed towards equilibrating cellular activity of water to that of the surrounding medium.

Animals↗

PubMed1984

Effects of Pb2+ added in vitro on Ca2+ movements in isolated mitochondria and slices of rat kidney cortex.

We have studied the effects of Pb2+ added in vitro on the movements of Ca2+ in renal cortical mitochondria and tissue slices. The isolated mitochondria rapidly accumulated 45Ca2+ at 25 degrees by a mechanism that was dependent on respiration and inhibited 96% by ruthenium red. A concentration of 10 microM Pb2+ inhibited the Ca2+ accumulation at least as effectively as did ruthenium red. About 20% of the Ca2+ accumulation persisted at 1 degrees with a similar sensitivity to inhibitors, including 60% inhibition by Pb2+. Similar results were obtained when the accumulation of Ca2+ at 25 degrees was measured by means of a calcium-sensitive electrode, Pb2+ inhibiting by 80%. Calcium that had been accumulated by mitochondria at 25 degrees was released completely by the ionophore A23187 or by 10 microM Pb2+. The release induced by Pb2+ was greatly inhibited by ruthenium red. The Ca2+ content of tissue slices of renal cortex increased 4-fold during incubation at 1 degree while the Ca2+ content of mitochondria within the slices more than doubled, the latter being determined by isolation of mitochondria from the slices after incubation. The presence of Pb2+ (200 microM) in the incubation medium of the slices substantially reduced the entry of Ca2+ into the whole slices and into mitochondria within the slices. When the slices preincubated at 1 degree were warmed to 25 degrees in oxygenated medium, they brought about a net extrusion of Ca2+, some of which was derived from the mitochondria; Pb2+ did not alter the final level of Ca2+ then attained in the slices, but it caused a significant decrease in the quantity retained in the mitochondria. We conclude that Pb2+ both inhibits the uptake of Ca2+ by renal cortical mitochondria and displaces Ca2+ from them, these effects occurring whether the mitochondria are isolated or in situ.

Animals↗

PubMed1988

Uptake of ochratoxin A by slices of pig kidney cortex.

Ochratoxin A (OCT A) is a nephrotoxin causing selective necrosis of the proximal tubule. Being an organic anion OCT A might be expected to enter the tubule cells by the organic anion transport system. Pig renal cortical slices were used to characterize the OCT A transport. OCT A (5 X 10(-3) mM) was accumulated against a concentration gradient with a slice to medium ratio of 8.9 +/- 2.9 in the presence of oxygen. This accumulation was markedly reduced in a nitrogen atmosphere (S/M ratio = 2.9 +/- 0.5). OCT A accumulation was dependent on medium concentration. With increasing concentration (5 X 10(-4)-5 X 10(-1) mM), slice to medium ratio initially rose from 6.9 +/- 2.0 to 11.7 +/- 1.2 whereupon it declined to 5.4 +/- 1.1. This pattern indicates that both carrier mediated transport and intracellular metabolism may contribute to the net accumulation of the toxin. OCT A (10(-4) to 1 mM) inhibited p-aminohippurate (PAH) and phenolsulphophthalein (PSP) uptake in a dose-dependent manner. Up to 10(-1) mM, OCT A did not inhibit acetylation of PAH suggesting that aerobic metabolism and the energy supply for the transport process were unaffected. Kinetic studies revealed a competitive inhibition of the PSP transport. It is concluded that OCT A enters the proximal tubule cells by the common organic anion transport system.

Animals↗

PubMed1992

Reconstitution and partial purification of calcium transport activity from rat kidney cortex.

An ATP-dependent Ca2+ uptake system from rat renal cortical basolateral membranes was solubilized with Triton X-100 and reconstituted into liposomes with lecithin. In the presence of Mg2+, Ca2+ uptake in the reconstituted vesicles was time and ATP dependent and was inhibited by vanadate. Ca2+ uptake in basolateral membrane vesicles depleted of endogenous calmodulin was enhanced by exogenous calmodulin and depressed by R-24571. This sensitivity to calmodulin and R-24571 was lost upon reconstitution in the presence and absence of leupeptin. Vesicles containing Ca2+ uptake activity were separated by gradient centrifugation after Ca2+ was taken up and accumulated as calcium phosphate in the vesicles. This resulted in Ca2+ uptake activity that was enriched 25 times. However, Ca(2+)-dependent adenosinetriphosphatase (ATPase) activity was not enriched significantly. This Ca(2+)-ATPase had two kinetic forms for Ca2+: one was a high-affinity low-capacity form; the other had a low affinity and high capacity. The Ca(2+)-ATPase activity also had two kinetic forms for ATP. All kinetic forms were inhibited by Mg2+. Vanadate, calmodulin, and R-24571 had no effects on Ca(2+)-ATPase activity. A protein doublet of Ca(2+)-dependent hydroxylamine-sensitive phosphorylated intermediates was demonstrated at 125 and 136 kDa in the purified vesicles. This doublet was not altered by addition of leupeptin throughout the purification.

Adenosine Triphosphate↗

PubMed1983

Different isotonic density gradients in separation of renin granules from rat kidney cortex.

Crude renin granule preparations isolated from the rat renal cortex were further purified in isotonic conditions (300 mOsm/kg) using various density gradient materials. It was not possible to separate renin granules from other subcellular organelles using dextran, 40,000-sucrose or metrizamide-sucrose gradients at about 300 mOsm/kg. When osmolality of dextran-sucrose gradients was increased, some separation was found but both renin granules and mitochondria gained density. During a short centrifugation (4640 X g, 30 min) renin granules remained intact and appeared in two populations in Percoll-sucrose gradients. The apparently heavier (larger) particles (at 1.12-1.13 kg/l) were greatly purified from mitochondria (80 X purification vs. the whole homogenate), protein (120 X) and lysosomes (24 X). Electron micrographs demonstrated many dense core granules. The fraction containing apparently lighter (small) granules (at 1.08-1.09 kg/l) was heavily contaminated with mitochondria and lysosomes. During longer centrifugation (4640 X g, 60 min), only one major peak showing renin activity was observed at 1.12-1.13 kg/l, and other cell organelles were lighter. Hence the two renin populations evidently do not differ in density but rather in size. In the animals kept on a low-sodium diet, both types of renin granules were increased.

Animals↗

PubMed1996

The effects of administering quinacrine during ultraprofound hypothermia on warm ischemic kidney cortex tissue.

Recent advances have led to increased use of ultraprofound hypothermia for cardiopulmonary bypass, organ preservation and trauma patients and have introduced the possibility of targeted pharmacologic intervention during the hypothermic period. In this study, rabbit renal cortex slices were used to examine the effect of administering quinacrine (100 microM) during hypothermia induced after a warm ischemic injury (60 min at 37 degrees C) on recovery of biochemical function during 3.5 hr of simulated warm reperfusion. In ischemic tissue slices, ATP content was reduced to near zero and only recovered about 50% by the end of reperfusion. Hypothermic storage of ischemic slices for 18 hr restored slice ATP content to about 80% of control levels but was followed by a decline during reperfusion to levels similar to ischemic slices. Administering quinacrine (100 microM) during 18 hr of hypothermic storage of ischemic slices resulted in a significant and sustained increase in slice ATP content during warm reperfusion. Slices stored at hypothermia only 3 hr with quinacrine had reduced swelling during reperfusion even though total ATP content was unaffected. Administering quinacrine (100 microM) only during reperfusion after ischemia or hypothermia did not affect tissue ATP content. This study showed that drug administration during hypothermic storage has potential therapeutic benefits for resuscitating tissues after warm ischemia and is more effective than the same drug given only during reperfusion. Tissue pretreatment was not required to obtain improved function in this study which suggests that future adaptations of these principles may have practical applications for specific clinical conditions where ischemic and reperfusion injury are significant factors.

Adenosine Triphosphate↗

PubMed1985

Gentamicin binding to brush border and basolateral membranes isolated from rat kidney cortex.

The interaction of gentamicin with renal cortex of rats has been studied in vitro by means of a binding assay to brush border and basolateral membranes. Gentamicin specifically bound to plasma membrane fractions, compared to other subcellular fractions. Gentamicin binding to brush border and basolateral membranes was markedly inhibited by polycations such as spermine, and was slightly inhibited by high concentrations of tetraethylammonium. The treatment of phospholipase A2 to both types of membranes increased gentamicin binding, although the treatments by proteolytic enzymes and sulfhydryl reagent did not affect the binding. Gentamicin binding was increased in the brush border membranes treated with acidic phospholipids, whereas it was decreased in the membranes treated with calcium. Judging from the determination of membrane surface charge by metachromasy of cationic dye, basolateral membranes seemed to contain more anionic sites than brush border membranes. The alterations of gentamicin binding described above correlated with the changes of anionic charge on the membranes, indicating a charge interaction between gentamicin and anionic binding sites on the membranes. The addition of other aminoglycoside antibiotics to the incubation mixture induced significant reductions in the binding of gentamicin in the order of aminoglycosides according to their positive charge. The present results suggest that the characteristics of gentamicin binding to brush border and to basolateral membranes are essentially similar, and therefore the renal accumulation of gentamicin may be regulated by the transport of gentamicin across both plasma membranes.

Aminoglycosides↗

PubMed1998

Cadmium inhibits vacuolar H(+)-ATPase and endocytosis in rat kidney cortex.

The mechanism of cadmium (Cd)-induced damage in the mammalian proximal tubule that is manifested by defects in reabsorption of various compounds, is poorly understood. A vacuolar H(+)-ATPase (V-ATPase) in proximal tubule (PT) brush border and intracellular vesicles may be affected by Cd, and this may influence intracellular vesicle trafficking and reabsorption of the filtered proteins. We studied the effects of Cd on V-ATPase and endocytosis in rat renal PT in vivo and on acidification mechanisms in isolated renal cortical organelles in vitro. The V-ATPase activity in brush border membrane (BBM) from Cd-intoxicated rats was 40% lower compared to that in control animals. Immunofluorescence studies in cortical tissue sections and Western blot studies in BBM from Cd-treated rats showed a strongly decreased abundance of the 31 kDa and 70 kDa V-ATPase subunits. Functional studies in vivo showed a dramatically diminished endocytosis of fluorescein-labeled dextran in PT cells from Cd-treated animals, whereas morphological studies revealed a loss of endocytic invaginations and subapical vesicles in the same cells. In studies in vitro, Cd inhibited V-ATPase activity in a concentration- and time-dependent manner in both BBM and endocytic vesicles, whereas in endocytic vesicles, Cd inhibited ATP-driven intravesicular acidification and accelerated the dissipation of transmembrane pH gradients. We conclude that Cd may impair acidification in cell organelles by (a) causing a loss of V-ATPase protein in their limiting membranes, (b) inhibiting the intrinsic V-ATPase activity, and (c) dissipating the transmembrane pH gradient. This may inhibit endocytosis of filtered proteins and impair vesicle-mediated recycling of some membrane transporters, thus contributing to the loss of reabsorptive capacity of the PT.

Acid-Base Equilibrium↗

PubMed1988

Cisplatin-induced lipid peroxidation and decrease of gluconeogenesis in rat kidney cortex: different effects of antioxidants and radical scavengers.

The present in vitro study was performed to investigate the effect of the nephrotoxic anticancer agent cisplatin (CP) on lipid peroxidation, on pyruvate-stimulated gluconeogenesis and on p-aminohippurate (PAH) accumulation in rat renal cortical slices. In addition, the inhibitory effects of the antioxidants and radical scavengers N,N'-diphenyl-p-phenylenediamine (DPPD), (+)-cyanidanol-3 or alpha-tocopherol on CP-induced lipid peroxidation and CP-induced decrease of gluconeogenesis and the inhibitory effect of DPPD on CP-induced decrease of PAH accumulation were evaluated. Slices were incubated in a CP-containing medium for different periods of time (7.5-300 min) and at different concentrations (0.025-1.5 mg/ml). Lipid peroxidation was monitored by measuring the production of malondialdehyde (MDA). Accumulation of PAH was expressed as slice to medium concentration ratio. Pyruvate-stimulated gluconeogenesis, measured as glucose production, was determined after a subsequent 60- or 15-min incubation in a pyruvate-containing, CP-free medium. CP led to a time- and concentration-dependent increase in MDA production, a time- and concentration-dependent decrease of pyruvate-stimulated gluconeogenesis and a time-dependent decrease of PAH accumulation in renal cortical slices. Decrease of gluconeogenesis preceded MDA production and decrease of PAH accumulation. Antioxidants reduced CP-induced MDA production and CP-induced decrease of accumulation of PAH, but did not reverse CP-induced decrease of gluconeogenesis. This might indicate, that the generation of free radicals and subsequent lipid peroxidation may play a role, at least in part, in inducing CP nephrotoxicity. There could be more than one mechanism of CP-induced nephrotoxicity, since decrease of gluconeogenesis preceded MDA production and decrease of PAH accumulation and could not be inhibited by antioxidants and radical scavengers.

Animals↗

PubMed1981

Further studies on properties of renin granules isolated from rat kidney cortex.

The properties of renin granules isolated from rat renal cortex were studied. Renin granules were thermolabile since in 10 min at 0 degrees C twice as much renin was released as at +37 degrees C. Addition of Ca++ (10(-6) M - 10(-2) M) did not affect the spontaneous release at +37 degrees C, pH 6.5, during 10 or 30 min incubation. However, when pH was elevated to above 7, renin release was significantly increased by Ca++ (10(-3) M). Additions of various amounts of KCl, NaCl or MgCl2, which increased the osmolality less than 20 mOsm/kg, did not affect the stability of the renin granules. Mg-ATP (0.5 and 5 mM) as well as Mg-GTP (5 mM) stabilized renin granules at +37 degrees C, pH 6.5, but the corresponding nitrogen analogues Mg-AMP-PNP and Mg-GMP-PNP (0.5 and 5 mM) were not effective. Neither did Mg-AMP (5 mM) nor ATP (5 mM) without Mg++ affect the renin release. No stabilization was observed by Mg-ATP and Mg-GTP in the purified granule preparations. The results suggest the importance of the cleavage of the terminal phosphate in the stabilization process. When the granules prepared at 300 mOsm/kg were first kept at hyperosmotic medium (range 300-1650 mOsm/kg) and then moved back to 300 mOsm/kg, the granules tend to lyse the more the greater was the reduction of the osmolality. The granules were more stable in isotonic sucrose than in isotonic ionic medium.

Calcium Chloride↗

PubMed2005

Ontogeny and effects of thyroid hormone on beta1-adrenergic receptor mRNA expression in ovine fetal kidney cortex.

OBJECTIVE: Previous studies indicate that thyroidectomy (TX) decreases renin gene expression in ovine fetal renal cortex in late gestation. Fetal ovine renin-containing renocortical cells become increasingly responsive to beta-adrenergic stimulation as gestation proceeds. Increases in plasma thyroid hormone concentrations parallel this change, suggesting that there is a positive developmental relationship between the two. To examine this hypothesis, we determined the ontogeny of beta1-adrenergic receptor (beta1R) mRNA expression, and the effect of thyroid hormone on in vivo and in vitro expression in fetal sheep. METHODS: Renocortical tissue was obtained from naive, TX, and sham-operated fetuses to determine beta1R mRNA levels. Renin-containing renocortical cells from TX or sham fetuses were treated with isoproterenol (Iso) or forskolin (FSK) for analysis of cellular cyclic adenosine monophosphate (cAMP) levels. Renocortical cells from naive fetuses were treated with triiodothyronine (T3) to assess cellular beta1R mRNA levels. Fetal plasma thyroxine (T4) level was determined. RESULTS: Renocortical beta1R mRNA expression increased significantly between 100 and 140 days' gestational age (dGA), while TX attenuated this increase (P <.01). Renocortical cellular cAMP levels were higher in sham compared to TX fetuses following incubation with Iso or FSK (P <.05). Cells incubated with T3 exhibited significantly increased beta1R mRNA expression (P <.05). CONCLUSION: The data suggest that thyroid hormone may be involved in modulating ovine fetal renocortical beta1R gene expression during development. We speculate that the increased beta1R mRNA expression in renal cortical cells as development progresses may mediate the increases in renin gene response to beta-adrenergic stimulation in late gestation.

Adrenergic beta-Agonists↗

PubMed1975

Interaction of parathyroid hormone with membranes of kidney cortex: degradation of the hormone and activation of adenylate cyclase.

Plasma membranes prepared from rat renal cortex contain both a parathyroid hormone-sensitive adenylate cyclase and a potent proteolytic activity which degrades the hormone into peptide fragments. The degree and pattern of degradation was determined by subjecting incubation mixtures to gel filtration and ion exchange chromatography. Estimation of the degree of degradation by acid precipitation of the intact hormone was inadequate since metabolism of the hormone apparently generated acid-insoluble fragments. When parathyroid hormone was incubated with membrane fraction, the capacity of its stimulatory effect on adenylate cyclase decreased steadily. This decrease of PTH activitiy could be closely related to the degradation of intact hormone by the same membrane preparation. The adenylate cyclase and degradative activity appeared to exist in similar membrane structures since they could not be separated by centrifugation through sucrose density gradients. The degradation of the hormone could not be inhibited by Trasylol and pancreatic or soybean trypsin inhibitors and was only slightly inhibited by ribonuclease and benzamidine. Histone (1 mg per ml), on the other hand, was able to decrease the degradation of the hormone and prevent the loss of its activity. Radioimmunoassay of the incubation mixtures showed that the rapid degradation of both amino- and carboxy-terminal regions of the hormone was prevented by histone. The oxidized, inactive hormone was also degraded to the same extent by the renal cortical membrane. Furthermore, the degradative activity was also found in plasma membrane preparations of renal medulla and liver. This lack of hormone and tissue specificity suggests that similar degradative activity exists in all tissues and that caution should be exercised in estimating hormonal potency based on activation of adenylate cyclase.

Adenylyl Cyclases↗