A Discussion on Triggered Enzyme Systems in Blood Plasma || A History and Review of the Kinin System

A History and Review of the Kinin SystemAuthor(s): Ashley MilesSource: Proceedings of the Royal Society of London. Series B, Biological Sciences, Vol. 173, No.1032, A Discussion on Triggered Enzyme Systems in Blood Plasma (Jul. 1, 1969), pp. 341-349Published by: The Royal SocietyStable URL: http://www.jstor.org/stable/75847 .

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Proc. Roy. Soc. B. 173, 341-349 (1969)

Printed in Great Britain

D. THE KININ SYSTEM

A history and review of the kinin system

BY SIR ASHLEY MILES, SEC.R.S.

The Lister Institute of Preventive Medicine, London, S.W. 1

INTRODUCTION

The first two plasma systems, clotting and fibrinolysis, we have so far dealt with in this meeting emerged largely from an analysis of clinically obvious end-results. The kinin system differs in two respects: first, it emerged largely from studies of

laboratory artifacts which suggested that a number of pharmacologically active substances could be derived from plasma; and secondly, in that the biological significance of its end results, whether physiological or pathological, is in most instances still a matter for conjecture.

This pharmacological odyssey has been concerned with studies of four biological effects. Two were on blood vessels, namely vasodilatation, which when induced

systemically is manifested as hypotension, and increased permeability of the walls of capillaries and venules to plasma proteins. The third effect is contraction of certain kinds of smooth muscle which, since the rat uterus is the most commonly used for in vitro tests, we may call uterotonic; and the fourth as Professor Keele and his colleagues first demonstrated, is the production of pain. All these four effects

may be induced by the end product of the plasma system-the kinins. As with

many other biological systems, the haphazard discovery of the different elements of the kinin system has engendered an almost equally haphazard growth of termin-

ology, and there is not yet a generally agreed rationalization of the terms in use. But at least the term 'kinin' is general enough not to offend the claims either of

priority or logic; and covers a group of unbranched polypeptides containing some nine to eleven amino acid residues. Kinins are formed by enzyme action on proteins, and we may with equally inoffensive generality call the substrate from which

they are split off 'kininogens', and the enzymes which do the splitting, 'kinino-

genases'. Kinin systems are not peculiar to plasma. In man, kininogenases are found in

urine and in the secretions of glands like the pancreas and the salivary gland; and their origin, nature and fate appear to be different from those of the plasma kininogenases (see Werle & Marier 1952; Moriya, Pierce & Webster I963; Webster & Pierce 1963). Moreover, both the glandular and the plasma kininogenases may have their counterparts in a number of other animals, like the guinea-pig, rat, ox, pig, horse, and certain birds (see, for example, Erdos, Back, Sicuteri & Wilde 1966). I shall confine myself to the human plasma system, merely noting that some of the

[ 341 ]



342 Sir Ashley Miles (Discussion Meeting)

pioneer steps in the definition of the kinin system were taken with glandular systems in man, and with both glandular and plasma systems in a variety of animals.

HISTORY

Hypotensive effects The concepts of the kinin system has been slow in growing (table i a). As regard

hypotensive effects, about the end of the first decade of the century both alkaline

(Vaughan & Wheeler 1907) and acid (Siegfried 90o6) hydrolysates of proteins were known to produce hypotensive shock in laboratory animals and, what is of

particular interest as an example of a triggered system, hypotensive substances were known to arise when fresh serum was treated with suspensions of relatively large-particulate foreign matter, like kaolin (Ritz & Sachs 191 ), or with antigen- antibody precipitates (Friedemann I909). In retrospect, the requirement of fresh serum for eliciting the effect of antigen-antibody complexes can be seen as the need for material in which the system had not already been activated and discharged, or degraded by mild heating or storage; at the time, however, it was taken as an indication that complement was involved (Friedberger i9io). At this time too an association was established between acute anaphylactic shock, in which hypotension is a cardinal sign, and the appearance in plasma of proteases, and of 'serotoxins' whose toxicity is in part attributable to the action of the proteases.

TABLE 1. GROWTH OF THE CONCEPT OF THE PLASMA KININ SYSTEM

(a) hypotensive effects (b) vascular permeability effects

1906-7 products of in vitro proteolysis 1909 anaphylactic 'serotoxins' 1909 anaphylactic 'serotoxins' 1938 leucotaxine 1910 fresh serum treated with antigen- 1953 bradykinin

antibody complexes 1955 PF 'proteases' 1911 fresh serum treated with kaolin 1960 plasmin and plasma kallikrein 1933 plasma kallikrein 1949 bradykinin 1955 PF 'proteases'

The subject was advanced more decisively in I933 by Kraut, Frey & Werle

(i933), who described a hypotensive substance in extracts of pancreas which they named 'kallikrein'. Pancreatic kallikrein was shown to release a smooth muscle contractor when added to plasma as substrate (Werle, Gotze & Keppler I937). Werle and his colleagues also described a kallikrein in plasma, which could be activated by treatment of the plasma with acid or with substances like chloroform or acetone (Werle & Berek 1948). In Werle's terminology the plasma system con- sisted of kallikrein, which when activated attacked a protein substrate 'kalli-

dinogen', releasing a vasoactive and uterotonic principle, 'kallidin'. In 1949 Rocha e Silva (Rocha e Silva, Beraldo & Rosenfeld 1949) described a

smooth muscle contractor released by trypsin from preparations of plasma pseudo globulin that proved also to be hypotensive, which he named 'bradykinin'.



History and review of the kinin system

Finally, at the Lister Institute we found in the plasma of the guinea-pig (Wilhelm, Miles & Mackay I955), rat (Wilhelm I956), rabbit (Wilhelm, Mill, Sparrow, Mackay & Miles I958) and man (Mill, Elder, Miles & Wilhelm 1958) apparently enzymic, globulin permeability factors (symbolized by PF) that were also hypotensive (see Mason & Sparrow 1965). From their susceptibility to trypsin inhibitors the factors appeared to be proteolytic enzymes.

Permeability effects There is a similar convergence in the history of permeability factors (table 1 b).

The tissue oedema appearing in anaphylactic shock was associated, and thought to be significantly so, with protease activation in the blood (Jobling & Petersen I914; see also Ungar & Hayashi I958). Leucotaxine, a permeability factor found

by Menkin (I938) in 24 h old inflammatory exudates, appeared to be a polypeptide, and was assumed to result from proteolysis in vivo; and later material from the in vitro hydrolysis of proteins with a similar biological action was found to be a mixture of polypeptides, which were most active as permeability factors when the amino acid residues numbered between 8 and 14 (Duthie & Chain 1938; Spector I95I). I have already mentioned the globulin permeability factor (PF) in connexion with hypotensive effects; crude preparations of bradykinin were shown in 1953 (see Rocha e Silva 1964) to increase vascular permeability; and later two more globulins, plasmin and plasma kallikrein (Miles & Wilhelm I960) were added to the list of plasma permeability factors.

STRUCTURE OF PLASMA KININS

It remains to add that the structure of Rocha e Silva's bradykinin was established both by analytic (Elliott, Lewis & Horton I960) and synthetic (Boissonas, Gutt- mann & Jaquenoud I960) methods, and that of Werle's kallidin by analytic methods (Pierce & Webster 196 ). Bradykinin is a nonapeptide, with the structure

Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg bradykinin

Lys-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg kallidin

Met-Lys-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg another plasma kinin

FIGURE 1. The amino acid sequence of three plasma kinins.

shown in figure 1, and kallidin a decapeptide, lysyl-bradykinin. These two kinins are products of the plasma kinin system per se; a third, methionyl-lysyl-bradykinin (Habermann i966) is formed from plasma kininogen by, e.g. the action of non-plasma proteases. In the pure state kinins are highly active. For example, concentrations of bradykinin as low as 10-1? are sufficient to stimulate the rat uterus in vitro (Elliott, Horton & Lewis I960).

343



Sir Ashley Miles (Discussion Meeting)

ACTIVATION OF THE SYSTEM

Werle released kinins by the acidification of plasma, or by shaking plasma with chloroform, thereby activating kininogenases. These are relatively brutal methods; a much more gentle method-and therefore perhaps more nearly related to natural

processes-came to light with the discovery in 1954 by Armstrong, Keele and their

colleagues (Armstrong, Keele, Jepson & Stewart 1954) that exposure of plasma to

glass led to the transient appearance of a substance, later identified as a kinin,

60

/ 10

exposure (min) 30/ /

lo- 60

5- 30 exposure to ,, _ \t tX^/~~~~~ \glass (min)

o, ) ~ J "O 10 30 90 270 810

reciprocal of plasma dilution (logarithmic scale)

FIGURE 2. Glass-activation of permeability increasing substances in graded dilutions of human plasma.

which caused pain and stimulated smooth muscle. In the following year, at the Lister Institute, we described permeability factors activated by letting dilute serum stand for an hour or so; and accordingly called it PF(dil) (Miles & Wilhelm

1955). But an analysis of the course of this activation, which was carried out in

glass tubes, showed that permeability factors first appeared transiently in un-

diluted plasma, and more slowly and more permanently, in dilute; and it became clear first, that activation was glass-dependent and second, that the production of PF(dil) after 1 h was optimal in dilute plasma because a natural inhibitor of the

PF, quick acting in undiluted plasma, was at 1/100 diluted to in effective concentrations (figure 2). Since the affix 'dil' is therefore misleading, I shall refer

344




below to this substance simply as PF. Dilution of plasma in glass also induces the formation of uterotonic kinins.

It was established by Margolis (1958) that glass activation depended on the

adsorption to glass surfaces of a 'contact factor' from the plasma, which in turn activated kininogenases. Margolis identified the contract factor in human plasma as factor XII or Hageman factor, and showed that the kinin system of Hageman- trait plasma was not capable of being glass activated (Margolis I959). Substances

analogous to Hageman factor operate in the plasma kinin systems of the guinea- pig and rat. It is to be expected that preparations of Hageman factor and of

kininogenases, on injection into certain animals, will trigger off the endogenous kinin systems and thus increase vascular permeability; because the animals have unactivated plasma kininogenases and kininogens in the blood; and because both the intercellular fluid of vascularized tissue and the lymph, in containing repre- sentatives of the plasma proteins, also contain the components of the kinin system.

100- --\\II k a

xi Y

- / o 50-

_ x

-0 0255 2 4 5 6\ 0 0-25 0-5 1 2 3 4 5 6

period of exposure to glass (h)

FIGURE 3. Glass-activation of 1/100 human plasma. k, kinin, indicating kininogenase activation; p, permeability increasing substances; a, substances that activate kininogenases in intact plasma.

The appearance of permeability factors during in vitro activation cannot, however, be explained solely in terms of factor XII, kininogenases and kinins.

During a continuing exposure of plasma to glass a permeability factor remains after kinins and kininogenases are no longer detectable (Mason & Miles I962; Miles I964). In figure 3, which illustrates part of an extensive analysis of the phenomenon by my colleague, Miss Brenda Mason, it is evident that when dilute plasma is exposed to glass for periods of up to 6 h a PF appears that

persists long after the disappearance of kininogenases and kinins. The corres-

ponding substance in plasma fractions is both permeability-increasing and

hypotensive. The persistent PF activates intact plasma in the absence of

glass; but, as Miss Mason (to be published) has established, in several decisive

respects, including susceptibility to di-isopropylfluorophosphonate, it is distinct not

345




only from the activating material adsorbed to glass-which she prefers to call 'surface' rather than 'contact' factor-but also from the plasma kininogenases, particularly in its relative heat stability and its inability to affect kininogens. It needs surface factor for its activation, and itself activates kininogenases in the absence of surface factor. There are also molecular distinctions to be made. For example, as Dr Becker and his colleagues showed, human kininogenase separates with the y-globulin fractions of plasma having a sedimentation constant of 11-3, and PF with S5-7 l-globulins (Kagen, Leddy & Becker I963).

SKETCH OF A KININ SYSTEM

We are now in a position to devise a scheme for the successive activation of a number of inert precursors in plasma, ending in kinin production. The scheme in figure 4 is set out on the same lines as Professor Macfarlane's (1964) orderly cascade of events in the clotting sequence. It should be emphasized that apart from the kinins, none of these components has been definitely characterized; isolation in a reasonably"active state, relatively free from other components, is the best that can be said of most of them. The nature of the Hageman factor I leave to the haematologists. On the assumption that P.F. and kininogenase are globular proteins, their sedimentation constants suggest molecular weights respectively of about 90000 and 350000; kininogenase, if a y-globulin, is therefore probably a dimer. Human kininogens are glycoproteins with a molecular weight of about 50000 (Pierce & Webster I966).

surface contact

proXIlI -- XII

pro-PF PF

prokininogenase--?" -kininogenase kininases

.inn. oI I. inactive kininogen -- kinm F peptides

FIGURE 4. A scheme of the plasma kinin system in man.

We have one major association with clotting and fibrinolysis, in that activation of factor XII can initiate all three processes. There is another association with fibrinolysis, to be discussed by Dr V. Eisen, in that plasmin can act as a kinino-

genase and perhaps as an activator of prokininogenase. We have no direct evidence that activation of pro-p.f. is a necessary link in the chain of events; it is certainly a possible link, and judging from the way pro-PF is exhausted during the activation of the kininogenase, it seems a very probable one. The only step definitely established as proteolytic is the splitting of kinins from kininogens; PF, however,

346




from its susceptibility to protease inhibitors, and, for what it is worth as evidence, its ability to split artificial amino acid esters, like TAMe, appears also to be a

protease. The scheme is, of course, grossly oversimplified. From the behaviour of plasma

after semi-selective exhaustion of various components of the system in plasma and of plasma fractionated in various ways, and from the effect of exogenous proteolytic enzymes on the system, several investigators from Werle onwards have idependently postulated more than one kininogenase, each with its appro- priately susceptible kininogen (Margolis & Bishop I963; Armstrong & Mills I964; Vogt I966; Eisen I966). The kinins usually produced are bradykinin and lysyl- bradykinin. Some investigators hold that the one or the other appears according to the nature of kininogenase concerned; others, that lysyl-bradykinin is formed and bradykinin derived from it by the action of a plasma aminopeptidase (Webster & Pierce I963). I have added the plasma kininases to the scheme. They are N-

carboxypeptidases and endopeptidases, the first removing the carboxy-terminal arginine from the kinins (Erdos, Renfrew, Sloane & Wohler 1963), and the second the terminal phenylalanine plus arginine, leaving biologically inactive peptides (Yang & Erd6s I967).

There is much else that could be added to figure 4. For example plasma contains inhibitors of PF (Wilhelm et al. I955) and ofkininogenases (Margolis 1959; Wrebster & Ratnoff I96I), apparently large-molecular, whose function in controlling activation is not yet clear. One of the inhibitors provides an association with the complement system, apparently being identical with the inhibitor of the esterase of the C'1 component of complement see Landerman, Webster, Becker & Ratcliffe I962; Donaldson & Evans i963). Preparations of the C'1 esterase, moreover, increase vascular permeability-though it is not known whether this effect is mediated by kinins (Ratnoff & Lepow :1963). It may be significant too of a connexion with

complement that after antigen-antibody reactions in vivo, or in suitably sensitized tissues in vitro, there is an activation of the kinin system (Beraldo I950; Brockle- hurst & Lahiri I960; see also E. L. Becker, this volume p. 383).

BIOLOGICAL SIGNIFICANCE OF THE SYSTEM

I propose to finish with some remarks on the biological significance of the kinin system, since I suppose we are not attempting to relate the various plasma system for purely intellectual satisfactions; but have some hope of practical issues in

physiology and pathology. In my introduction I implied that in no natural biological phenomenon had a

case been unequivocally made for the participation of the kinin system. I stress the word 'unequivocally'. There is plenty of evidence that triggering of the kinin system is associated with various physiological and pathological states. In table 2 some of these states are roughly classified. In many cases the argument for a causal linkage is highly plausible. The kinin system, as we have pieced it together, with

347




its inhibitors, kininases and so forth, is admirably adapted for controlled responses to triggering stimuli, and kinins certainly produce many of the effects observed in these various states. The readiness with which the system is activated, however, makes it difficult to judge whether activation is the cause or an accompaniment of the observed effects. Guilt by association, as the late Senator Joseph McCarthy so

amply and lamentably demonstrated, is no substitute for proof; and with the kinin

system, the main hope of a proof lies in the diminution of the naturally occurring effects by some form of selective depletion or inhibition.

TABLE 2. PHYSIOLOGICAL AND PATHOLOGICAL STATES IN MAN

ASSOCIATED WITH THE ACTIVATION OF PLASMA KININS

vasoactive permeability effects glandular secretion angioneurotic oedema carcinoid gouty arthritis shock, traumatic inflammation, traumatic

infective infective immunological immunological

pain

Nature, so far very lavish with variant human beings for the elucidation of the

clotting process, has obliged with selective depletion of bits of the kinin system only in two instances-Hageman-deficient man, and persons with hereditary angioneurotic oedema. Unfortunately, direct study of Mr Hageman and his like does not help us to determine whether kinins are responsible, for example, for

inflammation, because, as far as is known, Hageman-trait persons do not inflame

abnormally (Ratnoff & Miles 1964). Angioneurotic oedema is more informative, because liability to the periodic attacks of oedema is associated with an apparently hereditary deficiency of a single inhibitor of C'I esterase and PF and kininogenase, leaving apparently a kinin system which can be too readily activated (Becker &

Kagen I964). Convincingly selective depletion of the kinin system, whether by removal of

components or by their effective antagonism in vivo by exogenous inhibitors, has not yet proved possible. We still await the synthesis of highly specific, non-toxic

antagonists of one or more of the essential members of the plasma kinin systems, including the kinins, for tests of the physiological and pathological significance of the kinin system.

REFERENCES (Miles)

Armstrong, D., Keele, C. A., Jepson, J. B. & Stewart, J. W. I954 Nature, Lond. 174, 791. Armstrong, D. & Mills, G.L. I964 Biochem. Pharmac. 13, 1393. Becker, E. L. & Kagen, L. I964 Ann. N.Y. Acad. Sci. 116, 866. Beraldo, W. T. 1950 Am. J. Physiol. 163, 283.

Boissonas, R. A., Guttmann, S. & Jaquenoud, P. . 9A60 Helv. chim. Acta 43, 1349. Brocklehurst, W. E. & Lahiri, S. C. 1962 J. Physiol., Lond. 160, 15P. Donaldson, V. H. & Evans, R. R. 1963 Am. J. med. 35, 37. Duthie, E. S. & Chain, E. B. 1939 Br. J. exp. Path. 20, 417.

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History and review of the kinin system 349

Eisen, V. I966 J. Physiol., Lond. 186, 133P. Elliott, D. F., Horton, E. W. & Lewis, G. P. 1960 J. Physiol., Lond. 150, 6P. Elliott, D. F., Lewis, G. P. & Horton, E. W. 1960 Biochem. biophys. Res. Commun. 3, 87. Erd6s, E. G., Back, N., Sicuteri, F. & Wilde, A. F. (eds.) 1966 Hypotensive peptides. New

York: Springer-Verlag. Erd6s, E. G., Renfrew, A. G., Sloane, E. M. & Wohler, J. R. 1963 Ann. N.Y. Acad. Sci.

104, 222. Friedberger, E. 9I0o Z. ImmunForsch. exp. Ther. 4, 636. Friedemann, U. 1909 Z. ImmunForsch. exp. Ther. 2, 591. Habermann, E. 1966 Arch. exp. Path. Pharmak. 253, 474.

Jobling, J. W. & Petersen, W. 1914 J. exp. Med. 19, 480.

Kagen, L. J., Leddy, J. P. & Becker, E. L. I963 J. clin. Invest. 42, 1353. Kraut, H., Frey, E. K. & Werle, E. 1933 Hoppe-Seyler's Z. physiol. Chem. 222, 73. Landerman, N. S., Webster, M. E., Becker, E. L. & Ratclifie, H. E. 1962 J. Allergy 33, 330. Macfarlane, R. G. I964 Nature, Lond. 202, 495. Margolis, J. 1958 J. Physiol., Lond. 144, 1. Margolis, J. 1959 Aust. J. exp. Biol. med. Sci. 37, 329.

Margolis, J. & Bishop, E. I963 Aust. J. exp. Biol. med. Sci. 41, 293. Mason, B. & Miles, A. A. I962 Nature, Lond. 196, 587. Mason, B. & Sparrow, E. M. 1965 Br. J. Pharnac. Chemother. 25, 257. Menkin, V. 1938 J. exp. Med. 67, 129. Miles, A. A. 1964 Ann. N.Y. Acad. Sci. 116, 855. Miles, A. A. & Wilhelm, D. L. 1955 Br. J. exp. Path. 36, 71. Miles, A. A. & Wilhelm, D. L. 1960 The biochemical response to injury, p. 51. Oxford:

Blackwell. Mill, P. J., Elder, J. M., Miles, A. A. & Wilhelm, D. L. 1958 Br. J. exp. Path. 39, 343.

Moriya, H., Pierce, J. V. & Webster, M. E. 1963 Ann. N.Y. Acad. Sci. 104, 172. Pierce, J. V. & Webster, M. E. 1961 Biochem. biophys. Res. Commun. 5, 353. Pierce, J. V. & Webster, M. E. 1966 See Erdos et al. 1966, p. 130. Ratnoff, O. D. & Lepow, I. H. 1963 J. exp. Med. 118, 681. Ratnoff, 0. D. & Miles, A.A. . 964 Br. J. exp. Path. 45, 328. Ritz, H. & Sachs, H. 1911 Berl. klin. Wschr. 48, 987. Rocha e Silva, M. I953 See Rocha e Silva, M. I964 in Injury, inflammation and immunity,

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Siegfried, M. 1906 Hoppe-Seyler's Z. Physiol. Chem. 48, 54.

Spector, W. G. 1951 J. Path. Bact. 74, 67. Ungar, G. & Hayashi, H. 1958 Ann. Allergy 16, 542. Vaughan, V. C. & Wheeler, S.M. 907 J. infect. Dis. 4, 476. Vogt, W. 1966 See Erd6s et al. 1966, p. 185. Webster, M. E. & Pierce, J. V. 1963 Ann. N.Y. Acad. Sci. 104, 91. Webster, M. E. & Ratnoff, O. D. 1961 Nature, Lond. 192, 180. Werle, E. & Berek, U. 1948 Angew. Chem. 60A, 53. Werle, E., Gotze, W. & Keppler, K. 1937 Biochem. Z. 289, 217. Werle, E. & Maier, L. 1952 Biochem. Z. 323, 279. Wilhelm, D. L. 1956 Proc. R. Soc. Med. 49, 575. Wilhelm, D. L., Miles, A. A. & Mackay, M. E. 1955 Br. J. exp. Path. 36, 82. Wilhelm, D. L., Mill, P. J., Sparrow, E. M., Mackay, M. E. & Miles, A. A. 1958 Br. J. exp.

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A Discussion on Triggered Enzyme Systems in Blood Plasma || A History and Review of the Kinin System

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