An Inactive, Prorenin-like Substance in Human Kidney and Plasma

1980 ◽  
Vol 59 (s6) ◽  
pp. 29s-33s ◽  
Author(s):  
S. A. Atlas ◽  
J. H. Laragh ◽  
Jean E. Sealey ◽  
T. E. Hesson
Keyword(s):  
Molecular Weight ◽  
Affinity Chromatography ◽  
Human Plasma ◽  
Concanavalin A ◽  
Binding Protein ◽  
Human Kidney ◽  
Renin Activity ◽  
Cibacron Blue ◽  
Active Renin ◽  
Inactive Renin

1. Plasma prorenin (inactive renin), which accounts for about 70% of the total renin in human plasma, was almost completely separated from active renin by affinity chromatography on Cibacron blue F3G-A-agarose. The slight residual renin activity present in the prorenin peak can be removed on concanavalin A-Sepharose, demonstrating that prorenin is completely inactive. 2. The renin activity of both human renal cortical extract and renal perfusate increased after incubation with trypsin. This trypsin-activable renin accounted for 15 and 40% of the total renin in extract and perfusate respectively. 3. Trypsin-activable renin from both renal extract and renal perfusate was, like plasma prorenin, almost completely separated from active renin on Cibacron blue F3G-A-agarose. After additional chromatographic steps, the trypsin-activable renin from renal cortical extract was found to be completely inactive. 4. We conclude that human kidney contains, and is able to release, a trypsin-activable renin that resembles plasma prorenin. It may differ from many of the 60 000 molecular-weight forms of renin previously identified in renal extracts, since these possess considerable intrinsic renin activity and probably represent a complex of renin with a binding protein.

Affinity chromatography of human plasma gelsolin with polyphosphate compounds on immobilized Cibacron Blue F3GA

1990 ◽  
Vol 526 ◽  
pp. 397-406 ◽  
Author(s):  
Hiroaki Ito ◽  
Hideo Yamamoto ◽  
Yoshihiro Kimura ◽  
Hiroshi Kambe ◽  
Toshikazu Okochi ◽  
...  
Keyword(s):  
Affinity Chromatography ◽  
Human Plasma ◽  
Cibacron Blue ◽  
Plasma Gelsolin ◽  
Cibacron Blue F3ga

Activators of Inactive Renin (‘Prorenin’) in Human Plasma: Their Connection with Kinin Formation, Coagulation and Fibrinolysis

1979 ◽  
Vol 57 (s5) ◽  
pp. 89s-92s ◽  
Author(s):  
F. H. M. Derkx ◽  
B. N. Bouma ◽  
H. L. Tan-Tjiong ◽  
M. A. D. H. Schalekamp
Keyword(s):  
Human Plasma ◽  
Hageman Factor ◽  
Active Renin ◽  
Normal Plasma ◽  
Inactive Renin ◽  
Fletcher Factor ◽  
Free Plasma ◽  
Renin Activation ◽  
Dependent Pathway ◽  
Proteolytic Pathways

1. Human plasma was treated at 4°C with acid, trypsin, plasmin, streptokinase, urokinase, active Hageman factor fragment (β-XIIa) and β-XIIa-activated plasma prekallikrein (Fletcher factor). The conversion of inactive into active renin (activation) was studied in normal plasma (n = 10), Hageman factor-deficient plasma (n = 2), Fletcher factor-deficient plasma (n = 1) and plasminogen-free plasma (n = 4). 2. In normal plasma inactive renin was activated at pH 7·5 after treatment at pH < 4·0; at pH 3·3 the results were the same as with trypsin. This was also the case in plasminogen-free plasma. In Hageman factor-deficient plasma and in Fletcher factor-deficient plasma, however, the quantities of renin that were activated after acidification were much smaller than with trypsin. The addition of physiological amounts of active kallikrein to pH 3·3-pretreated Hageman factor-deficient plasma caused complete activation of renin. In contrast, the addition of active Hageman factor fragment to pH 3·3-pretreated Fletcher factor-deficient plasma had little or no effect. 3. Plasmin, streptokinase-activated plasminogen and urokinase-activated plasminogen activated inactive renin in pH 4·0-pretreated normal plasma as well as in pH 4·0-pretreated Hageman factor-deficient plasma and Fletcher factor-deficient plasma. 4. It is concluded that inactive renin is activated by two separate proteolytic pathways: one pathway depends on both Hageman factor and plasma prekallikrein, and the other pathway depends on plasminogen. In the Hageman factor-dependent pathway plasma kallikrein and not Hageman factor is the major activator of inactive renin. It is assumed that pH 3·3-treatment of plasma destroys the major inhibitors of kallikrein and that pH 4·0-treatment destroys the major inhibitor of plasmin.

scholarly journals Actin affinity chromatography in the purification of human, avian and other mammalian plasma proteins binding vitamin D and its metabolites (Gc globulins)

1984 ◽  
Vol 218 (3) ◽  
pp. 805-810 ◽  
Author(s):  
J G Haddad ◽  
M A Kowalski ◽  
J W Sanger
Keyword(s):  
Vitamin D ◽  
Affinity Chromatography ◽  
Human Plasma ◽  
Plasma Protein ◽  
Binding Protein ◽  
Binding Activity ◽  
Single Step ◽  
Affinity Column ◽  
Vitamin D Metabolites ◽  
Molar Ratios

The human plasma protein binding vitamin D and its metabolites (Gc globulin; group-specific component) has been isolated from human plasma by column affinity chromatography on gels to which monomeric actin was covalently attached. Rabbit skeletal-muscle G-actin was covalently coupled to amino-agarose gels before the application of human plasma. At actin/protein molar ratios of 4-8:1, excellent recovery (approximately 58%) of purified binding protein was achieved. After 0.75 M-NaCl washes, the binding protein was eluted from the columns in 3 M-guanidinium chloride, dialysed and analysed. These eluates contained the binding protein as 34-100% of the total protein, reflecting a 130-fold average purification in this single step. In the presence of Ca2+, gelsolin (another plasma protein that binds actin) was apparently retained by the affinity column, but this was prevented by chelation of plasma Ca2+. The actin affinity step also was effective in the isolation of the binding protein from rat, rabbit and chicken plasma, as indicated by autoradiographs of purified fractions analysed by gel electrophoresis after incubation with 25-hydroxy[26,27-3H]cholecalciferol. Further isolation by hydroxyapatite chromatography yielded a purified binding protein which displayed characteristic binding activity toward vitamin D metabolites and G-actin, and retained its physicochemical features. This brief purification sequence is relatively simple and efficient, and should prove to be useful to investigators studying this interesting plasma protein.

scholarly journals Partial purification and characterization of chick-embryo prolyl 3-hydroxylase

1979 ◽  
Vol 183 (2) ◽  
pp. 303-307 ◽  
Author(s):  
K Tryggvason ◽  
K Majamaa ◽  
J Risteli ◽  
K I Kivirikko
Keyword(s):  
Molecular Weight ◽  
Chick Embryo ◽  
Ethylene Glycol ◽  
Affinity Chromatography ◽  
Concanavalin A ◽  
Gel Filtration ◽  
Partial Purification ◽  
Purification And Characterization ◽  
Chick Embryo Extract

Prolyl 3-hydroxylase was purified up to about 5000-fold from an (NH4)2SO4 fraction of chick-embryo extract by a procedure consisting of affinity chromatography on denatured collagen linked to agarose, elution with ethylene glycol and gel filtration. The molecular weight of the purified enzyme is about 160000 by gel filtration The enzyme is probably a glycoprotein, since (a) its activity is inhibited by concanavalin A, and (b) the enzyme is bound to columns of this lectin coupled to agarose and can be eluted with a buffer containing methyl alpha-D-mannoside. The Km values for Fe2+, 2-oxoglutarate, O2 and ascorbate in the prolyl 3-hydroxylase reaction were found to be very similar to those previously reported for these co-substrates in the prolyl 4-hydroxylase and lysyl hydroxylase reactions.

The Nature of Inactive Renin in Human Plasma and Amniotic Fluid

1978 ◽  
Vol 55 (1) ◽  
pp. 41-50 ◽  
Author(s):  
A. A. Shulkes ◽  
R. R. Gibson ◽  
S. L. Skinner
Keyword(s):  
Molecular Weight ◽  
Amniotic Fluid ◽  
Human Plasma ◽  
Gel Filtration ◽  
Dilution Effect ◽  
Exchange Chromatography ◽  
Acid Activation ◽  
Temperature Dependent ◽  
Inactive Renin ◽  
Full Activation

1. The properties of inactive and active renin in human plasma and amniotic fluid were studied chromatographically. Activation was achieved at pH 3.3 with and without added pepsin. 2. Acid activation of renin was time- and temperature-dependent but was inhibited by dilution of the sample. The dilution effect was corrected by adding pepsin. Such characteristics indicate that activation at low pH is catalysed by intrinsic enzymes. 3. Separation and/or dilution of the activating enzyme during ion-exchange chromatography concealed the eluted position of inactive renin and reduced the amount recovered. Only after full activation of the eluted renin was achieved with added pepsin was a distinct peak of inactive renin exposed. 4. At pH 7.5 inactive renin carried a lower negative charge than the active enzyme. This charge difference was lost after activation. 5. No molecular-weight differences between active, inactive renin or the International Renin Standard were detected by gel filtration. No renin of larger molecular weight was present. 6. These findings will be helpful in purification studies of human inactive renin.

scholarly journals Rabbit erythrocyte band 3: a receptor for staphylococcal alpha toxin

1980 ◽  
Vol 26 (4) ◽  
pp. 524-531 ◽  
Author(s):  
Indar Maharaj ◽  
Hugh B. Fackrell
Keyword(s):  
Molecular Weight ◽  
Staphylococcus Aureus ◽  
Affinity Chromatography ◽  
Concanavalin A ◽  
Hemolytic Activity ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Band 3 ◽  
Alpha Toxin ◽  
Rabbit Erythrocyte

Enzymes known to specifically cleave the band 3 component of the rabbit erythrocyte membrane were found to reduce both the hemolytic sensitivity to and the binding of the alpha toxin of Staphylococcus aureus. Lectins which bind to band 3 also inhibited the toxin. Lectins which do not bind to band 3 have no effect. Purified band 3, isolated by affinity chromatography on a concanavalin A column, was homogeneous by polyacrylamide gel electrophoresis, had a molecular weight of 100 000, and inhibited the hemolytic activity of alpha toxin. Antibodies to the toxin–toxoid receptor were serologically indistinguishable from antiband 3.

Two Different Types of Inactive Renin in Human Plasma: Molecular and Kinetic Properties

1984 ◽  
Vol 67 (4) ◽  
pp. 421-425
Author(s):  
Yoichi Imamura ◽  
Kunio Hiwada ◽  
Tatsuo Kokubu
Keyword(s):  
Molecular Weight ◽  
Human Plasma ◽  
Kinetic Properties ◽  
Carbohydrate Composition ◽  
Ph Profile ◽  
Isoelectric Points ◽  
Different Types ◽  
Km Value ◽  
Inactive Renin ◽  
Fresh Plasma

1. We separated inactive renin in human plasma into two types, adsorbed and non-adsorbed, by chromatography on a concanavalin A-Sepharose column. About 75% of fresh plasma inactive renin was adsorbed to the column, and the rest passed through it. Non-adsorbed and adsorbed inactive renins were partially purified. 2. Non-adsorbed inactive renin had a molecular weight of 48000 and an isoelectric point of 5.44. Adsorbed inactive renin had a molecular weight of 46000 and isoelectric points of 5.56 and 5.80. 3. After activation with trypsin, both activated inactive renins were similar with respect to molecular weight (45000), thermostability, Km value (0.56 μmol/l) and pH profile. But pI values of both activated inactive renins differed. 4. These results indicate that there exist in human plasma two different types of inactive renin which differ in carbohydrate composition.

The relationship between renin isoelectric forms and renin glycoforms

1994 ◽  
Vol 267 (1) ◽  
pp. R244-R252
Author(s):  
S. A. Katz ◽  
J. A. Opsahl ◽  
P. A. Abraham ◽  
M. J. Gardner
Keyword(s):  
Sialic Acid ◽  
Affinity Chromatography ◽  
Isoelectric Focusing ◽  
Concanavalin A ◽  
Plasma Clearance ◽  
Rat Kidney ◽  
Clearance Rates ◽  
Con A ◽  
Active Renin ◽  
The Relationship

Active renin can be separated into multiple isoelectric forms using shallow gradient isoelectric focusing and into multiple glycoforms using concanavalin A (Con A) affinity chromatography. The relationship between renin isoelectric forms and glycoforms has not been previously determined. In this study, each of three renin Con A glycoforms from rat kidney was composed of significantly different proportions of six renin isoelectric forms; glycoforms with the greatest affinity for Con A contained proportionally less of the acidic isoelectric forms than those with the least affinity for Con A. A set of compartmental models accurately predicted previously measured differential plasma clearance rates of the three renin glycoforms based on their corresponding isoelectric form proportions. We conclude that 1) each Con A renin glycoform is composed of significantly different proportions of isoelectric forms, and 2) the different proportions of isoelectric forms found in Con A glycoforms are sufficient to account for the differential renin plasma clearance rates demonstrated previously for renin glycoforms in the rat. These data suggest that the isoelectric and glycoform heterogeneity of active renin are, in fact, closely related and may result from variable and interrelated mannose (Con A affinity) and sialic acid (charge) attachments to renin.

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