Identification of Renin and Renin-like Enzymes in Rat Brain by a Renin-Specific Antibody

1980 ◽  
Vol 59 (s6) ◽  
pp. 45s-47s ◽  
Author(s):  
V. J. Dzau ◽  
Amy Brenner ◽  
Neremiah Emmett ◽  
E. Haber
Keyword(s):  
Affinity Chromatography ◽  
Rat Brain ◽  
Specific Antibody ◽  
Angiotensin I ◽  
Ph Optimum ◽  
Brain Extracts ◽  
Antibody Inhibition ◽  
Neutral Proteases

1. The major angiotensin I-generating activity of rat brain extracts has a pH optimum different from that of renal renin and is not inhibited by renin specific antibody. 2. Affinity chromatography utilizing renin specific antibody, pepstatin and α-casein yielded fractions that resembled renal renin more closely with respect to antibody inhibition and pH optimum as well as an absence of the ability to hydrolyse haemoglobin. 3. We conclude that rat brain contains a host of enzymes with angiotensin I-generating activity including acid and neutral proteases and an enzyme with the immunochemical identity of renal renin. The biosynthetic origins of this renin-like protein remain uncertain.

Evidence for renin in rat brain: differentiation from other reninlike enzymes

1982 ◽  
Vol 242 (5) ◽  
pp. E292-E297 ◽  
Author(s):  
V. J. Dzau ◽  
A. Brenner ◽  
N. L. Emmett
Keyword(s):  
Rat Brain ◽  
Protease Activity ◽  
Lithium Bromide ◽  
Acid Protease ◽  
Brain Extract ◽  
Affinity Column ◽  
Angiotensin I ◽  
Ph Optimum ◽  
The Brain ◽  
Neutral Proteases

We observed that unfractionated rat brain extract incubated with substrate at pH 6.0 yielded 12 times the quantity of angiotensin I as incubations at pH 7.4, but the enzyme activity measured at pH 6 was not primarily due to renin. To examine the existence of renin in brain, we used three methods of affinity chromatography (pepstatin-, renin-specific antibody-, and alpha-casein-Sepharose) to fractionate the angiotensin I-generating enzymes in the brain. 1) Brain extract applied to renin-specific column eluted a peak of angiotensin-releasing activity (ARA) that had a pH optimum of 6.0. This ARA was inhibited by antirenin antibody. Another peak of ARA with a pH optimum of 4 appeared in the nonbound fraction. This peak was not affected by antirenin antibody and had acid protease activity. 2) Pepstatin affinity column elution with lithium bromide yielded an early ARA peak (pH optimum 6.5), inhibited by antirenin antibody and a later peak (pH optimum 4.0) not inhibited by antirenin antibody. The latter contained acid protease activity. 3) alpha-Casein-Sepharose column also separated neutral proteases and immunoreactive renin from acid protease capable of generating angiotensin. In summary, rat brain contains a host of angiotensin I-generating enzymes that can be detected and separated as neutral and acid proteases and immunoreactive renin depending on the pH of the assay and conditions of purification. These findings indicate the presence of an enzyme with immunoidentity to renin in rat brain but do not imply local biosynthesis.

Definitive Evidence for Renin in Rat Brain by Affinity Chromatographic Separation from Protease

1978 ◽  
Vol 55 (s4) ◽  
pp. 121s-123s ◽  
Author(s):  
Tadashi Inagami ◽  
Hideyoshi Yokosawa ◽  
Shigehisa Hirose
Keyword(s):  
Affinity Chromatography ◽  
Rat Brain ◽  
Chromatographic Separation ◽  
Protease Activity ◽  
Brain Extract ◽  
Angiotensin I ◽  
Definitive Evidence

1. Angiotensin I-generating activity of rat brain extract was separated into two components by affinity chromatography on a casein—Sepharose gel column. 2. The component without affinity to the gel was identified as true renin on the basis of its sensitivity to anti-renin antibody and the lack of protease activity. 3. The second renin-like component with affinity to the gel was a protease insensitive to the anti-renin antibody. Its renin-like activity examined with sheep substrate was pronounced compared with the rate of angiotensin I generation from the rat substrate. 4. It was concluded that rat brain contains true renin, which can be detected by the use of rat substrate but can be masked when examined with sheep substrate.

Actin purification from a gel of rat brain extracts

Biochimie ◽  
1984 ◽  
Vol 66 (7-8) ◽  
pp. 531-537 ◽  
Author(s):  
Nicole Levilliers ◽  
Monique Peron-Renner ◽  
Gérard Coffe ◽  
Julio Pudles
Keyword(s):  
Rat Brain ◽  
Brain Extracts

scholarly journals The diphosphoinositide kinase of rat brain

1968 ◽  
Vol 106 (4) ◽  
pp. 791-801 ◽  
Author(s):  
M. Kai ◽  
J. G. Salway ◽  
J. N. Hawthorne
Keyword(s):  
Rat Brain ◽  
Kinase Activity ◽  
Ion Concentration ◽  
Supernatant Fraction ◽  
Thiol Groups ◽  
Ph Optimum ◽  
Adult Rat ◽  
Phosphatidylinositol Kinase ◽  
Adult Rat Brain ◽  
Ammonium Sulphate Fractionation

1. The supernatant fraction of adult rat brain contains a diphosphoinositide kinase. 2. Formation of triphosphoinositide by the enzyme in the presence of ATP and Mg2+ ions was shown with labelled ATP or labelled diphosphoinositide. 3. The kinase was also activated by Ca2+, Mn2+ and Co2+ ions, but to a smaller extent than by Mg2+ ions. 4. In the presence of optimum Mg2+ ion concentration the enzyme was inhibited by Ca2+ ions. 5. Activity did not depend on thiol groups and the pH optimum was 7·3. 6. The dialysed supernatant fraction had no diglyceride kinase activity and negligible phosphatidylinositol kinase activity. 7. Triphosphoinositide phosphomonoesterase was present but showed little activity under the conditions used to assay the kinase. 8. Diphosphoinositide kinase was purified by ammonium sulphate fractionation, ethanol treatment and chromatography on Sephadex G-200. 9. This purification removed much of the triphosphoinositide phosphomonoesterase.

The identification of acetyl-l-carnitylcholine in rat brain extracts and the comparison of its cholinomimetic properties with acetylcholine

1970 ◽  
Vol 48 (10) ◽  
pp. 709-722 ◽  
Author(s):  
E. A. Hosein ◽  
A. Kato ◽  
E. Vine ◽  
A. M. Hill
Keyword(s):  
Guinea Pig ◽  
Rat Brain ◽  
Total Activity ◽  
Guinea Pig Ileum ◽  
Molar Basis ◽  
Choline Esters ◽  
Brain Extracts

Acetyl-l-carnitylcholine (l-ACCh) was identified in rat brain extracts on paper chromatograms developed in butanol–water for 138 h. l-ACCh was also identified in brain extracts fractionated on t.l.c. plates and on Sephadex G-10 columns. In every instance l-ACCh was separated from the acetylcholine (ACh) present and the ACh-like activity of l-ACCh was about 20% of the total activity in the extract. Both l-ACCh and ACh were found to be inseparable in a variety of chromatographic systems including electrophoresis. Treatment of these choline esters with cholinesterases showed that while true acetylcholinesterase hydrolyzed both l-ACCh and ACh, pseudocholinesterase destroyed only ACh. On a molar basis, the ACh-like activity of ACCh is one-half that of ACh on both the guinea pig ileum and frog rectus preparations. Like ACh, the ratio of the nicotinic to muscarinic potency of l-ACCh is unity. Mixtures of l-ACCh and ACh show summation of ACh-like activity on both the guinea pig ileum and frog rectus preparations.

Immunocytochemical and Biochemical Characterization of Angiotensin I and II in Cultured Neuronal and Glial Cells from Rat Brain

1988 ◽  
Vol 47 (2) ◽  
pp. 125-132 ◽  
Author(s):  
Klaus Hermann ◽  
Mohan K. Raizada ◽  
Colin Sumners ◽  
M. Ian Phillips
Keyword(s):  
Rat Brain ◽  
Glial Cells ◽  
Biochemical Characterization ◽  
Angiotensin I

scholarly journals Detection of active proteasome structures in brain extracts: proteasome features of August rat brain with violations in monoamine metabolism

Oncotarget ◽  
2017 ◽  
Vol 8 (41) ◽  
pp. 70941-70957 ◽  
Author(s):  
Pavel A. Erokhov ◽  
Yulia V. Lyupina ◽  
Alexandra S. Radchenko ◽  
Anna A. Kolacheva ◽  
Yulia O. Nikishina ◽  
...  
Keyword(s):  
Rat Brain ◽  
Monoamine Metabolism ◽  
Brain Extracts

ESTIMATION OF MARSUPIAL RENIN USING MARSUPIAL RENIN-SUBSTRATE

1972 ◽  
Vol 53 (1) ◽  
pp. 125-130 ◽  
Author(s):  
PAMELA A. SIMPSON ◽  
J. R. BLAIR-WEST
Keyword(s):  
Substrate Concentration ◽  
Structural Difference ◽  
Plasma Renin ◽  
Angiotensin I ◽  
Ph Optimum ◽  
Ph Profile ◽  
Renin Substrate ◽  
Grey Kangaroo ◽  
Rate Of Reaction ◽  
Highly Correlated

SUMMARY Bilateral nephrectomy of an Eastern Grey kangaroo (Macropus giganteus) increased plasma renin-substrate concentration approximately tenfold when compared with intact kangaroos. A preparation made from this plasma had a renin-substrate concentration of 3000 ng/ml. A pH profile of rate of reaction with pig renin had an optimum at pH 5·39. By comparison, the pH optimum of sheep renin-substrate was pH 6·15. Estimates of plasma renin concentration for kangaroos, wombats and wallabies, using kangaroo renin-substrate or sheep renin-substrate were highly correlated. Results from incubation with sheep renin-substrate were greater and hence indicate the advantage in using this substrate for marsupial renin estimation. The consistently large difference between sheep and kangaroo renin-substrate when incubated with renin from marsupial and eutherian species appears to be due to a structural difference between the two substrates, probably near the C-terminal end of the angiotensin I molecule.

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