The purification and characterization of multiple forms of mouse submaxillary gland renin

Ho, Siu-Cheong; Izumi, Hiroshi; Michelakis, Andrew M.

doi:10.1016/0304-4165(82)90281-1

Cited by 10 publications

(6 citation statements)

References 28 publications

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“…Purified mouse kidney renin was prepared by pepstatin-affinity, immunoaffinity, and CM-cellulose chromatographic techniques (16). The purity of this protein was judged by a single protein band in polyacrylamide gel electrophoresis.…”

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confidence: 99%

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Multiple Defects in the Renin-Angiotensin System in Alloxan-Diabetic Kidney

Ho¹,

Michelakis²

1985

Experimental Biology and Medicine

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A defect in the renin-angiotensin system has been shown in diabetic patients and experimental animals, in particular with nephropathy or autonomic neuropathy. The mechanism for this low plasma renin activity (PRA) is poorly understood. In order to clarify this defect, the renin-angiotensin system was studied in alloxan-induced diabetic and age-match control mice. In diabetic animals, kidney renin activity (KRA) was significantly lower than that of the controls, while plasma renin substrate (PRS) concentration was slightly higher and PRA was normal. The amount of injected radiolabeled renin extracted by the kidney was normal, but the amount extracted by the liver was significantly decreased in diabetic animals. On the other hand, the degradation of the extracted renin by both the kidney and the liver was elevated as compared to the controls. This high degradation rate was accompanied by a slight increase in lysosomal protease activity in the kidneys. In in vivo studies, isoproterenol-induced PRA was 20-fold in control animals. In diabetics, isoproterenol-induced PRA was attenuated and rose only four-to fivefold over basal level. The angiotensin converting enzyme (ACE) activity in the kidney was significantly decreased in the diabetic state. It is concluded that there were multiple defects in the renin-angiotensin system in this diabetic model, namely, a depletion of renin storage with subsequent loss of maximal responsiveness to the adrenergic agonist in renin release, an elevation of intrarenal renin degradation together with a deficiency in ACE which would possibly lead to a decrease in intrarenal formation of angiotensin 11. o 1985 Society for Experimental Biology and Medicine.

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confidence: 99%

“…PRA was determined in the presence of exogenous renin substrate (plasma from rats nephrectomized 48 hr previously) by 10-min incubation at 37°C to assure linear generation of angiotensin I (2). The angiotensin I generated was determined by radioimmunoassay ( 16). Kidney renin activity (KRA) was determined as described previously (2).…”

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confidence: 99%

Multiple Defects in the Renin-Angiotensin System in Alloxan-Diabetic Kidney

Ho¹,

Michelakis²

1985

Experimental Biology and Medicine

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“…For Ren-2 renin reacting with rat aogen at pH 7.4, it has been calculated that 1 nmol of the pure active enzyme is equivalent 16 GU of hog standard renin. 20,21 This calculation included a molecular weight (MW) of 40 000 Da, which, though corroborated, 22 is now identified as closer to 36 000 Da for nonglycosylated SMG Ren-2 renin. 18,23 Rat renin MW, which is glycosylated, has been reported to be 36 000 to 42 000 Da.…”

Section: Purified Reninsmentioning

confidence: 76%

Hypertension in the (mRen-2)27 Rat Is Not Explained by Enhanced Kinetics of Transgenic Ren-2 Renin

2003

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Abstract-Enhanced efficiency of the reaction between transgenic Ren-2 mouse renin and endogenous rat angiotensinogen has been suggested as 1 mechanism that contributes to the accelerated hypertension and increased tissue angiotensin of the (mRen-2)27 transgenic rat. This was tested in a study conducted at pH 7.4 in vitro that compared the kinetic constants of purified mouse Ren-2 and rat renin (each at 100, 75, 50, and 25 pmol/L) reacting with physiologic concentrations of rat angiotensinogen (0 to 4 mol/L). Under these conditions, the kinetic constants for Ren-2 (K m , 1.8 mol/L; K cat , 0.07/s; and K cat /K m , 0.04 L · mol Ϫ1 · s Ϫ1 ) were not different from rat renin. However, Ren-2 renin acting on its homologous mouse angiotensinogen was confirmed as being much slower. We conclude that hypertension in the Ren-2 rat is not related to renin kinetics. Other mechanisms are considered, with reference to human essential hypertension. Key Words: renin-angiotensin system Ⅲ angiotensinogen Ⅲ blood pressure Ⅲ hypertension, essential T he hypertensive (mRen-2)27 transgenic rat (Ren-2 rat) developed by Mullins et al 1 displays strong expression of the mouse Ren-2 transgene in several extrarenal tissues, 2 suppressed endogenous renal renin, and predominantly mouse inactive prorenin in the circulation secreted from extrarenal sources. [3][4][5] Most evidence indicates that it is the active component of mouse Ren-2 renin in tissues and plasma, reacting with the endogenous rat angiotensinogen (aogen), that induces and sustains the hypertension through conventional angiotensin II (Ang II)-dependent mechanisms, in particular because the hypertension is readily controlled by inhibition of the renin-angiotensin system (RAS). 6 However, this concept is at odds with the initial published evidence of suppressed or normal active plasma renin and angiotensin peptides in the adult Ren-2 rat, 1,7,8 reports that encouraged a search for additional hypertensive mechanisms. This transgenic rat strain has now been used widely to study various conditions relating to activation of the tissue RAS, including angiogenesis, cytokine activation, and profibrotic and inflammatory pathologic states, making it important to understand fully the fundamental processes inducing the severe hypertension.That plasma Ang II is not suppressed is now apparent, with resting levels increased up to 4-fold in both young 9 and adult 10 Ren-2 rats. One would expect a similar elevation of plasma active renin, but this depends on the pH at which the activity is estimated, 11,12 because mouse and rat renins have different pH optima when acting on rat aogen (pH 8.5 and 6.5, respectively). 5,11,12 Measured at pH 7.4 the Ren-2 rat plasma renin reaction is a mixed activity, mainly owing to the mouse transgenic renin, 11 and increases in renin activity have been found under these physiologic conditions. The lower level of rat renin in plasma is consistent with its low but not absent level in the kidneys. 11,12 By comparison with other high-renin models that cause malignant...

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“…The labeling of antibodies with z25I was carried out with chloramine T following the procedure described by Ho et al (17). Free n5I was removed by passing the labeled material over a column (3.0 × 0.5 cm) of Dowex AGI×8 (Bio-Rad Laboratories, Richmond, CA).…”

Section: Seed Soybean Agglu Tinin and A N Tibody Reagentsmentioning

confidence: 99%

Endogenous lectins from cultured soybean cells: isolation of a protein immunologically cross-reactive with seed soybean agglutinin and analysis of its role in binding of Rhizobium japonicum.

Ho¹,

Malek-Hedayat²,

Wang³

et al. 1986

The Journal of Cell Biology

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Abstract. Incubation of Rhizobium japonicum with the cultured soybean cell line SB-1, originally derived from the roots of Glycine max, resulted in specific adhesion of the bacteria to the plant cells. This binding interaction appears to be mediated via carbohydrate recognition, since galactose can inhibit the heterotypic adhesion but glucose cannot. Affinity chromatography, on a Sepharose column derivatized with N-caproyl-galactosamine, of the supernatant fraction of a SB-1 cell suspension after enzymatic removal of cell wall yielded a single polypeptide (Mr ,030,000) on immunoblotting analysis with rabbit antibodies directed against seed soybean agglutinin. Fluorescently labeled rabbit anti-seed soybean agglutinin also yielded specific immunofluorescent staining on the cell wall and plasma membrane of the SB-1 cells. These results suggest that one likely candidate that may mediate the recognition between the Rhizobium and the soybean cells is the endogenously produced SB-1 lectin. This notion is supported by the observation that rabbit anti-seed soybean agglutinin blocked the Rhizobium-soybean cell adhesion, whereas control antibodies did not.

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The purification and characterization of multiple forms of mouse submaxillary gland renin

Cited by 10 publications

References 28 publications

Multiple Defects in the Renin-Angiotensin System in Alloxan-Diabetic Kidney

Multiple Defects in the Renin-Angiotensin System in Alloxan-Diabetic Kidney

Hypertension in the (mRen-2)27 Rat Is Not Explained by Enhanced Kinetics of Transgenic Ren-2 Renin

Endogenous lectins from cultured soybean cells: isolation of a protein immunologically cross-reactive with seed soybean agglutinin and analysis of its role in binding of Rhizobium japonicum.

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