Purification and partial characterization of rat brain acid proteinase (isorenin)

Hackenthal, E.; Hackenthal, Renate; Hilgenfeldt, Ulrich

doi:10.1016/0005-2744(78)90088-8

Cited by 71 publications

(27 citation statements)

References 24 publications

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“…Although the earlier observation of a renin-like enzyme within various extrarenal tissues (14) may have been mostly due to cathepsin D (EC 3.4.23.5) (15,16), we have demonstrated the presence ofspecific renin, which is inhibitable by antibodies to renin, in various tissues of hogs (17) and rats (18). A particularly high renin activity was found in the rat adrenal (18).…”

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confidence: 73%

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Markedly elevated specific renin levels in the adrenal in genetically hypertensive rats.

Naruse¹,

Inagami²

1982

Proc. Natl. Acad. Sci. U.S.A.

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The specific renin (EC 3.4.99.19) activity in the adrenal of spontaneously hypertensive rats was determined by a method that is capable of distinguishing renin from nonspecific renin-like activity ofproteases by using specific antibody to renin. The renin level in the adrenals of adult spontaneously hypertensive rats with established hypertension was found to be 6-8 times as high as that of the normotensive control Wistar-Kyoto strain. The large difference in the adrenal renin level was observed even in 3-wk-old rats in which hypertension had not yet developed. The adrenal renin level was increased by bilateral nephrectomy in both the hypertensive and normotensive strains. A larger quantity of renin was found in the adrenal cortex than in the medulla, and the difference between the hypertensive strain and the normotensive strain was more prominent in the cortex than in the medulla. These results suggest possible involvement of adrenal renin in the development and in the early maintenance phase of hypertension in this animal model of human essential hypertension by affecting the adrenocortical or adrenomedullary activity, or both.The etiology of hypertension in spontaneously hypertensive rats (SHR) (1) has been of great interest in view of its resemblance to essential hypertension in humans. Elevated levels of dopamine f3-hydroxylase (EC 1.14.17.1) activity in plasma (2), mesenteric vessels, and adrenals (3) and decreased levels of dopamine ,3-hydroxylase activity (3) and norepinephrin (4) in the noradrenergic regions of the brain observed before the development of hypertension indicate a significant role of neurogenic factors in the initiation ofthe hypertension ofthis animal model (5). The aberrations in the levels of these neurogenic substances are no more than 2-fold. On the other hand, normal or subnormal levels of plasma renin (EC 3.4.99.19) have indicated the lack of a causative role for the renin-angiotensin system in spontaneous hypertension (6)(7)(8)(9)(10).However, the recent finding that the angiotensin-converting enzyme inhibitor, captopril (D-3-mercapto-2-methylpropanoyl-L-proline), normalizes the blood pressure of patients with essential hypertension and that of the SHR (11, 12) suggests a possible role of the renin-angiotensin system in this type of essential hypertension. Because the plasma level of renin activity is not increased and because captopril is known to be taken up by tissues (13), such a renin-angiotensin system may exert its effect by its local function.Although the earlier observation of a renin-like enzyme within various extrarenal tissues (14) may have been mostly due to cathepsin D (EC 3.4.23.5) (15, 16), we have demonstrated the presence ofspecific renin, which is inhibitable by antibodies to renin, in various tissues of hogs (17) and rats (18). A particularly high renin activity was found in the rat adrenal (18). Therefore, in an attempt to assess the possible role of the tissue renin-angiotensin system in the peripheral system of spontaneous hypertension, we have comp...

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confidence: 73%

“…Angiotensin I-generating activities have been reported in the adrenal gland (14,27,28). However, these tissues contain large amounts of cathepsins, which show nonspecific renin-like activity (16). This nonspecific activity often accounts for a sizable proportion of the total angiotensin I-generating activity of tissue extract.…”

Section: Discussionmentioning

confidence: 99%

Markedly elevated specific renin levels in the adrenal in genetically hypertensive rats.

Naruse¹,

Inagami²

1982

Proc. Natl. Acad. Sci. U.S.A.

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“…This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact. tivity of cathepsin D (14,15), parallel experiments were performed with the extract preincubated with specific antibodies to pure rat (27) and mouse renin (28) at 1:10,000 dilutions for 16 hr at 40C. At this dilution the antisera were found to inhibit 10 ng of renin completely, but not cathepsin D. The pure rat and mouse renin used for producing the antibodies were shown to satisfy multiple criteria of purity (27,28).…”

Section: Methodsmentioning

confidence: 99%

“…In the central system, demonstration of specific renin in brain extracts by affinity chromatography has established that the reninlike activity (1,2) is indeed due to specific renin (13) rather than a nonspecific action ofcathepsin D (14,15 (16)(17)(18). However, this enzyme is present on the luminal aspect of the vascular endothelium throughout the body (19,20), and the existence of the enzyme in brain parenchyma has not been demonstrated unequivocally.…”

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confidence: 99%

Renin, angiotensins, and angiotensin-converting enzyme in neuroblastoma cells: evidence for intracellular formation of angiotensins.

Okamura¹,

Clemens²,

Inagami³

1981

Proc. Natl. Acad. Sci. U.S.A.

110

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The mechanism of formation of various peptide hormones in neuronal cells in the brain is not clear. The question of whether brain angiotensin II is formed by an extracellular mechanism as in the peripheral system or by an intracellular mechanism can be answered by using cloned cells in culture. We have screened several neuroblastoma cell lines of rat and mouse origin and found at least three cell lines that contain renin (EC 3.4.99.19), angiotensin-converting enzyme (dipeptidyl carboxypeptidase; peptidyldipeptide hydrolase, EC 3.4.15.1), and angiotensins I and II. This finding was interpreted to indicate that in these cells angiotensin formation takes place by an intracellular mechanism, in contrast to the extracellular mechanism well known to occur in plasma. This study also demonstrates the existence of viable and cloned cell lines that produce renin.Many polypeptide hormones originally discovered in peripheral tissues are also present in various regions of the brain. The nature and localization of the enzymes involved in the processing of prohormone to hormone in the brain have not been established. Angiotensin II, apotentvasoconstrictoroctapeptide, has been found in various regions of the brain (1-4). The presence of its receptor in the brain (5, 6) and multiple physiological responses elicited by centrally administered angiotensin II (7-12) indicate a direct effect of this hormone on neuronal function. Although the extracellular pathway of angiotensin formation in the circulation is well established, the intracellular localization of angiotensin II in the brain (1-4) suggests that the pathway in the central nervous system may be different from that of the periphery.The first step of angiotensin formation in the periphery is mediated by the circulating enzyme renin (EC 3.4.99.19). In the central system, demonstration of specific renin in brain extracts by affinity chromatography has established that the reninlike activity (1, 2) is indeed due to specific renin (13) Experimental results favoring an intracellular mechanism of angiotensin formation in these cloned cells were obtained. MATERIALS AND METHODSNeuroblastoma Cells. Established neuroblastoma cell lines used were B82, B103 (21), and RT4E4 (22) of rat origin, and Neuro-2a (23), NB41A3 (24), and N4TG1 (25) derived from mouse neuroblastoma C-1300. Cells, except for NB41A3 cells, were cultured in Dulbecco's modified Eagle's medium containing 12.5% horse serum and 2.5% fetal calf serum until confluency (4-10 days). NB41A3 cells were cultured in Ham's F-10 medium with additions of horse and fetal calf serum to the same concentrations. Twenty-four hours prior to harvesting the cells, media were changed to serum-free solutions. Rat neuroblastoma cells were detached from culture flasks with 1 mM EDTA, washed twice with the serum-free medium, suspended in water containing a mixture of 1 mM diisopropyl phosphorofluoridate (iPr2P-F), 1 mM Captopril (an angiotensin-converting enzyme inhibitor; Squibb, leupeptin at 5 jig/ml, and 5 mM EDTA and lysed by fiv...

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“…This work resolved the raging controversy as to whether "iso-renin" in dog brain reported by Ganten et al 49 is renin. Day and Reid 50 and Hackenthal et al 51 had published data indicating that it was the nonspecific action of cathepsin D. Use of the casein-Sepharose column clearly demonstrated that although the great majority of the iso-renin is due to the nonspecific action of cathepsin D, a small amount of specific renin exists in the brain. 48 This was another very important application of affinity purification of renin.…”

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confidence: 99%

Purification of renin and prorenin.

Inagami

1991

Hypertension

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Purification and partial characterization of rat brain acid proteinase (isorenin)

Cited by 71 publications

References 24 publications

Markedly elevated specific renin levels in the adrenal in genetically hypertensive rats.

Markedly elevated specific renin levels in the adrenal in genetically hypertensive rats.

Renin, angiotensins, and angiotensin-converting enzyme in neuroblastoma cells: evidence for intracellular formation of angiotensins.

Purification of renin and prorenin.

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