Marion E. Webster

Hereditary angioneurotic edema: II. Deficiency of inhibitor for serum globulin permeability factor and/or plasma kallikrein☆

Abstract Studies of a 33-year-old Caucasian woman with hereditary angioneurotic edema and her two sons, of whom only one is definitely affected with the disease, have revealed that the inborn biochemical lesion is likely an inherited deficiency of serum inhibitor to plasma kallikrein and/or serum globulin permeability factor. The patient showed a significantly greater bluing response to intradermal injections of diluted autologous serum or to plasma kallikrein than did the normal controls and this response, unlike that of the control subjects, continued to increase for a period of hours. By use of an enzymatic determination utilizing p-toluenesulfonyl-l-arginine methyl ester as the substrate, it is possible to demonstrate a deficiency in both patients to an inhibitor of plasma kallikrein. A variety of other known permeability factors have been excluded as the potential agent involved.

The nature of the kallidins released from human plasma by kallikreins and other enzymes.

The kallikreins are hypotensive enzymes of endogenous origin whose injection intravenously in very minute quantities causes a rapid lowering of the blood pressure of the dog. Urinary, pancreatic and serum kallikreins were originally thought to be one substance. Since the pharmacologically active material was found in large amounts in the pancreas, Kraut, Frey and Werlel in 1930 named it kallikrein from the Greek word for pancreas, kallikreas. However, more recent e v i d e n ~ e ~ ~ , has made it clear that kallikreins derived from different sources or different species vary in their response to proteolytic inhibitors or to antibody (TABLE 1).

Action of the Kallikreins on Synthetic Ester Substrates

Summary Evidence is presented which suggests that the kallikreins derived from hog pancreas, human pancreas, human urine and human plasma are capable of digesting those synthetic esters which contain arginine as the specific amino acid residue. Highly purified hog pancreatic kallikrein also digests acetyl-L-tyrosine ethyl ester. Arginine esters and benzoyl-L-arginine amide prevented both inhibition of human urinary and pancreatic kallikreins by human plasma and inhibition of human plasma kallikrein by diisopropyl-fluorophosphate. Acetone activates not only plasma kallikrein but another proteolytic enzyme(s) which attack arginine esters and are not inhibited by SBTI.

The Purification and Some Properties of Two Different Kallidinogens from Human Plasma

Although bovine kininogen has been highly purified (Habermann et al., 1963; Suzuki et al, 1965), very little has been published on the purification of human plasma kallidinogen (Webster and Pierce, 1963). For the isolation of native kininogens, mild conditions of separation should be used. Denaturing treatments such as heat and low pH should be avoided. In addition, the use of a glandular kallikrein would seem preferable to the use of trypsin in following kininogen activity. The former enzyme is more likely to require undenatured substrates since it has a higher substrate specificity than trypsin, whereas the latter enzyme generally works best on denatured or fragmented kininogens.

URATE CRYSTAL INDUCED INFLAMMATION IN THE RAT: EVIDENCE FOR THE COMBINED ACTIONS OF KININS, HISTAMINE AND COMPONENTS OF COMPLEMENT

The inflammation produced by subplantar injection of sodium urate micro-crystals in the hindpaw of the rat was used as a model of gouty arthritis. Kinins are probably involved since carboxypeptidase B or soybean trypsin inhibitor partially (35%) blocked the edema. Serotonin does not participate since the edema was not prevented by methysergide or cyproheptadine. Histamine is apparently involved since many antihistaminics also partially (40%) suppressed this edema. Two antihistaminics, promethazine and tri-pelennamine, inhibited (75%) the edema as effectively as colchicine by mechanisms as yet not established. Treatment of rats with a combination of soybean trypsin inhibitor and a specific antihistaminic, triprolidine, inhibited the urate edema by 58-66%, which suggested that still another mediator was involved. Complement depletion of the rats also partially (30%) inhibited the edema. The combination of soybean trypsin inhibitor, triprolidine and complement depletion almost completely (80%) suppressed the...

The estimation of the kallidins in blood and urine

Abstract The pharmacological properties of the kallidins and the location of specific kallikreins in most glandular tissue and in plasma suggest that local activation of this enzyme system may be involved in a variety of physiological and pathological conditions; i.e. blood flow regulation, inflammatory reactions, allergic phenomena, various kinds of shock, etc. 1 Direct determination of the polypeptide should provide a more sensitive means for the detection of activation of the kallikrein-kallidin system than measurement of the kallidinogen (bradykininogen) content of the plasma. 2, 3 This report describes a sensitive and specific method for the determination of the kallidins in blood and urine.

Synthesis of prekallikrein and metabolism of plasma kallikrein by perfused rat liver

Abstract When livers from normal rats were perfused in situ with oxygenated Tyrodes solution containing bovine serum albumin, both active kallikrein and prekallikrein could be found in the perfusates. Based on five liver perfusates the average rate of synthesis was 6.6 ± 2.0 mU · hr −1 · (g liver) −1 when N- benzoyl - l - prolyl - l - phenylalanyl - l - arginyl -p- nitroanilide was used as substrate, and it was calculated that within 30 hr each liver could synthesize 50 per cent of the prekallikrein content of rat plasma. The liver was also capable of rapidly inactivating rat and human plasma kallikreins and plasmin but not rat urinary kallikrein. The rates of clearance from perfusate were biphasic, giving an initial rapid clearance (2 min) succeeded by a slower phase that followed an exponential curve. These data clearly show that the exsanguinated and perfused rat liver is capable of synthesizing plasma prekallikrein and of metabolizing plasma kallikrein and plasmin.

Interaction of leukocytes and endotoxin with the plasmin and kinin systems.

Leukocytes can generate a substance that, when added to some partially purified human kininogen, is capable of forming kinins. The addition of endotoxin or polystyrene latex particles to the incubated leukocytes doubled the amount of kinin generated. Certain preparations of kininogen, however, failed to allow kinin formation by the leukocytes. No evidence could be found that an activator of prekallikrein or a kallikrein was present in the granulocyte preparations. However, the addition of highly purified plasminogen to inactive kininogen preparations restored their ability to generate kinins in the presence of leukocytes. All the kininogen preparations that allowed kinin formation when incubated with leukocytes contained plasminogen. These data suggest that a plasminogen activator is present on the leukocyte surface. This activator activates plasminogen to form plasmin which in turn acts on kininogen to release a kinin and thus provides a mechanism for the formation of kinins in inflammatory exudates and during endotoxemia.

Immunological properties of the kallikreins

Abstract Rabbits immunized with crude or partially purified human urinary and pancreatic kallikreins developed antibodies which inhibited the vasodilator activity of the kallikreins in dogs. The antigenic heterogeneity of the kallikreins was shown by the presence of multiple precipitin bands in agar gel. The kallikrein-antikallikrein band was tentatively identified with the use of purified kallikreins. Antibody to human urinary kallikrein showed cross reaction both in agar gel and by inhibition of the vasodilator activity with human pancreatic kallikrein but not with dog urinary or hog pancreatic kallikrein. The normal inhibitor in rabbit serum could be differentiated from antibody by its stability to acetone and to storage at pH 3.0. Unlike the normal inhibitor, anti-body was not competitively inhibited by p- toluenesulfonyl - l - arginine methyl ester (TAMe) and was a more potent inhibitor of the vasodilator activity than the TAMe activity of kallikrein.

Kallidin (lysylbradykinin), the kinin formed from horse plasma by horse urinary kallikrein

Abstract Horse urinary kallikrein when incubated with horse plasma formed kallidin (lysylbradykinin) from the kininogens in the plasma. Horse plasma, like human plasma, was found to contain an aminopeptidase capable of converting kallidin to bradykinin. No evidence, however, could be found that the plasma contained an aminopeptidase capable of converting Met-Lys-bradykinin to kallidin, thus eliminating the possibility that the kallikrein had released Met-Lys-bradykinin which was converted to kallidin during the 1–5 min incubations. The method used for identification of the kinins is rapid, gives a good recovery and requires small amounts of plasma and enzyme.