Clinica Chimica Acta, 210 (1992) 233-235 0 1992 Elsevier Science Publishers B.V. All rights reserved. 0009-8981/92/$05.00
233
CCA 05371
Letter to the Editor
Activity of cathepsin H,B and metalloproteinase the serum of patients with acute myocardial infarction
in
(Received 16 December 1991; revision received 2 July 1992; accepted 16 July 1992)
Dear Editor, Several markers of myocardial infarction have been described. These include aspartate transaminase (AST), lactate dehydrogenase (LDH), LDH-1 isoenzyme or hydroxy-butyl-dehydrogenase (HBDH), creatine kinase (CK), CK-MB isoenzyme, myosin and troponine [ 11. Recent observations suggest that a subcellular redistribution of lysosomal proteinases may play a role in the progression of intracellular damage [l] and release of lysosomal cathepsin D (EC 3.4.23.5) may be an indicator of myocardial cell death. An in vitro proteolytic effect of rabbit cardiac procathepsin D (proCD) has been demonstrated by Samarel and Lesch [2]. The intracellular proteolytic processing of proCD does not result solely from autocatalytic activation, but requires at least one other protease, perhaps cathepsin B. Cysteine proteinases (cathepsin H,B) (EC 3.4.22.16, EC 3.4.22.1) and metalloproteinase (MMP-7ase, EC 3.4.24.15) activities have not been investigated in patients suspected of having acute myocardial infarction. We have determined these enzyme activities in serum. Controls were 11 healthy people with the similar age as the patients were. Among 9 patients with acute myocardial infarction serum AST, cathepsin H,B and MMP-7ase activities were determined on 23 occasions. Cathepsin H,B and MMP-7ase activities were compared with the AST activity 1, 2 and 3 days after infarction. The activities of cathepsin H and B (CH, CB) as lysosomal cysteine proteinases were measured by the method of Kirschke et al. [3] and MMP-7ase activity was determined according to Sohar et al. [4] using succinyl-Ala-Ala-Pro-Phe-7-(4methyl)coumarinyl-amide as substrate. The liberated 7-amino-4-methyl-coumarine Correspondence to: Aranka L&lb, School, Szeged, Hungary.
Department of Paediatrics, A. Szent-Gyiirgyi University Medical
234
TABLE I Serum AST, cathepsin H,B and MMP-7ase in myocardial infarct patients (AMI)
1 f S.D. 2 zt S.D. 3 *S.D. Control
AST (Ufl)
Cath. H (mu/ml)
Cath. B (mu/ml)
MMP-7ase (mu/ml)
63.75 85.96 57.67 30.25 85.3 58.2
186.6 93.2 151.9 56.8 160.57 52.58
4.33 2.0 3.78 i.99 3.85 1.67
16.67 19.93 11.57 6.55 8.71 5.93
28.4
104.8
3.51
7.55
7.1
6.5
0.51
3.29
group n = I I
zizS.D. D-probe 1st day 2nd day 3rd day Correiation
P < 0.001 n.s.
n.s. ns. n.s.
1st day 0.111 n.s. 0.111 n.s. 0.004 n.s.
2nd day 0.66 ns 0.19 ns 0.32 n.s
3rd day 0.26 ns. 0.45 n.s. 0.31 n.s.
coef#cients
between AST and Cath. H MMP-7 ase Cath. B r
P < 0.0.5
ns. ns. n.s.
was measured ~uorimetrically (Ex.: 360 nm, Em.: 460 nm) using a Hitachi 650-10 (Tokyo, Japan) spectrofluorimeter. Substrates were supplied by Enzymes System Products (Liver-more, CA). The correlations between the activities of AST and cathepsin H,B and MMP-7ase were investigated using linear correlation coefficients. CH activities were raised si~i~~a~tly on the first and third days. The enzyme activities were highest on the first day after heart attack and fell on the second and third day. The activity of cathepsin B and the MMP-7ase were not raised signilicantly, because of the high standard derivations. The investigated enzyme activities did not show any correlation with AST activity (Table I). Three of the patients died, of these two had extremely elevated CH, CB and MMP-7ase activities. It shows that the very high protease activity may indicate a severe infarct, but this should be proved in the future. Our data are correlated with results of Wildenthal and Decker [I]. They published that lysosomal proteases (CD), acid phosphatase and glu~osamidase are released from heart muscle after coronary ligation. We found that other types of lysosomal proteinases (cysteine proteinase CH, CB) were also released. The cysteine proteinases are activated in reduced state [5]. In our hypothesis lysosomal cysteine proteinases (CH, CB) could be activated in the heart during ishemia first time and they can activate the aspartic proteinases (CD) by limited proteolysis 161.Aspartic proteinase directly and via damage of inhibitors of cysteine proteinase (cystatins)
235
can again activate cysteine proteinases. This high activity of proteinases can be excreted into the blood through the damaged cell membranes. Aranka
LBsz1~5~,Istvan Sohirb, Istvan S4gic, Judit Kov;icsb and Attila Kov6csd
aDepartment of Paediotrics, ‘Department of Biochemistry, CIntensive Therap~t~c Care Unit and dl. Department of Internai medicine* A. Szent-Gyiirgyi University Medical &hoof, Szeged (Htatgary~
References 1
2
3 4
5
Wildenth~ K, Decker RS, Lysosomal enzyme distribution in normal and ischemic my~ardium. J Mol Cell Cardiol 1984;16(suppl. 1):14. Samarel A, Lesch M. In vitro proteolysis of rabbit cardiac procathepsin D. Program and Abstracts of the Meeting of the American Section of the International Society for Heart Research, Oklahoma City, USA, 13-15 Sept 1984. J Mol Cell Cardiol 1984;16(suppl. 1):32. Kirschkke H, Wood L, Roisen FJ, Bird JWC. Activity of iysosomal cysteine proteinase during differentiation of rat skeletal muscles. Biochem J ~983;21~871-877. Sohlr I, Fekete E, Yorke F, Cosentino B, Roisen FJ, Bird JWC. Proteinase activities in normal and dystrophic chicken myoblasts in culture. Intracell Prot Catab 198.5;629-63I. Sohar I, Katona G. Regulation of proteinase activities in mammalian tissues. Biol Chem HoppeSeyler 1992;in press.
Further Suggested Reading Hors&h M, KugIer Zs, Dibusz J, Falukiizi J. Nehany megfigyelts Diagnostics Pasteur Miozin IRMA kittel. Laborat Diagnosztika XVII/3, 1990;188(in Hungarian). Tischler ME, Fagan JM, Allen DK. Reduction-oxidation state as a mediator of hormonal control of muscle proteolysis. J Mol Cell Cardiol 1984;16(suppl. 1):36. Stein W. Laboratory diagnosis of acute myocardial infarction. Darmstadt: GIT Verlag, 1988. Kooistra T, Miilard PC, Lloyd JB. Role of thiols in degradation of proteins by cathepsins. B&hem J f 982;204:47I-477.
233
CCA 05371
Letter to the Editor
Activity of cathepsin H,B and metalloproteinase the serum of patients with acute myocardial infarction
in
(Received 16 December 1991; revision received 2 July 1992; accepted 16 July 1992)
Dear Editor, Several markers of myocardial infarction have been described. These include aspartate transaminase (AST), lactate dehydrogenase (LDH), LDH-1 isoenzyme or hydroxy-butyl-dehydrogenase (HBDH), creatine kinase (CK), CK-MB isoenzyme, myosin and troponine [ 11. Recent observations suggest that a subcellular redistribution of lysosomal proteinases may play a role in the progression of intracellular damage [l] and release of lysosomal cathepsin D (EC 3.4.23.5) may be an indicator of myocardial cell death. An in vitro proteolytic effect of rabbit cardiac procathepsin D (proCD) has been demonstrated by Samarel and Lesch [2]. The intracellular proteolytic processing of proCD does not result solely from autocatalytic activation, but requires at least one other protease, perhaps cathepsin B. Cysteine proteinases (cathepsin H,B) (EC 3.4.22.16, EC 3.4.22.1) and metalloproteinase (MMP-7ase, EC 3.4.24.15) activities have not been investigated in patients suspected of having acute myocardial infarction. We have determined these enzyme activities in serum. Controls were 11 healthy people with the similar age as the patients were. Among 9 patients with acute myocardial infarction serum AST, cathepsin H,B and MMP-7ase activities were determined on 23 occasions. Cathepsin H,B and MMP-7ase activities were compared with the AST activity 1, 2 and 3 days after infarction. The activities of cathepsin H and B (CH, CB) as lysosomal cysteine proteinases were measured by the method of Kirschke et al. [3] and MMP-7ase activity was determined according to Sohar et al. [4] using succinyl-Ala-Ala-Pro-Phe-7-(4methyl)coumarinyl-amide as substrate. The liberated 7-amino-4-methyl-coumarine Correspondence to: Aranka L&lb, School, Szeged, Hungary.
Department of Paediatrics, A. Szent-Gyiirgyi University Medical
234
TABLE I Serum AST, cathepsin H,B and MMP-7ase in myocardial infarct patients (AMI)
1 f S.D. 2 zt S.D. 3 *S.D. Control
AST (Ufl)
Cath. H (mu/ml)
Cath. B (mu/ml)
MMP-7ase (mu/ml)
63.75 85.96 57.67 30.25 85.3 58.2
186.6 93.2 151.9 56.8 160.57 52.58
4.33 2.0 3.78 i.99 3.85 1.67
16.67 19.93 11.57 6.55 8.71 5.93
28.4
104.8
3.51
7.55
7.1
6.5
0.51
3.29
group n = I I
zizS.D. D-probe 1st day 2nd day 3rd day Correiation
P < 0.001 n.s.
n.s. ns. n.s.
1st day 0.111 n.s. 0.111 n.s. 0.004 n.s.
2nd day 0.66 ns 0.19 ns 0.32 n.s
3rd day 0.26 ns. 0.45 n.s. 0.31 n.s.
coef#cients
between AST and Cath. H MMP-7 ase Cath. B r
P < 0.0.5
ns. ns. n.s.
was measured ~uorimetrically (Ex.: 360 nm, Em.: 460 nm) using a Hitachi 650-10 (Tokyo, Japan) spectrofluorimeter. Substrates were supplied by Enzymes System Products (Liver-more, CA). The correlations between the activities of AST and cathepsin H,B and MMP-7ase were investigated using linear correlation coefficients. CH activities were raised si~i~~a~tly on the first and third days. The enzyme activities were highest on the first day after heart attack and fell on the second and third day. The activity of cathepsin B and the MMP-7ase were not raised signilicantly, because of the high standard derivations. The investigated enzyme activities did not show any correlation with AST activity (Table I). Three of the patients died, of these two had extremely elevated CH, CB and MMP-7ase activities. It shows that the very high protease activity may indicate a severe infarct, but this should be proved in the future. Our data are correlated with results of Wildenthal and Decker [I]. They published that lysosomal proteases (CD), acid phosphatase and glu~osamidase are released from heart muscle after coronary ligation. We found that other types of lysosomal proteinases (cysteine proteinase CH, CB) were also released. The cysteine proteinases are activated in reduced state [5]. In our hypothesis lysosomal cysteine proteinases (CH, CB) could be activated in the heart during ishemia first time and they can activate the aspartic proteinases (CD) by limited proteolysis 161.Aspartic proteinase directly and via damage of inhibitors of cysteine proteinase (cystatins)
235
can again activate cysteine proteinases. This high activity of proteinases can be excreted into the blood through the damaged cell membranes. Aranka
LBsz1~5~,Istvan Sohirb, Istvan S4gic, Judit Kov;icsb and Attila Kov6csd
aDepartment of Paediotrics, ‘Department of Biochemistry, CIntensive Therap~t~c Care Unit and dl. Department of Internai medicine* A. Szent-Gyiirgyi University Medical &hoof, Szeged (Htatgary~
References 1
2
3 4
5
Wildenth~ K, Decker RS, Lysosomal enzyme distribution in normal and ischemic my~ardium. J Mol Cell Cardiol 1984;16(suppl. 1):14. Samarel A, Lesch M. In vitro proteolysis of rabbit cardiac procathepsin D. Program and Abstracts of the Meeting of the American Section of the International Society for Heart Research, Oklahoma City, USA, 13-15 Sept 1984. J Mol Cell Cardiol 1984;16(suppl. 1):32. Kirschkke H, Wood L, Roisen FJ, Bird JWC. Activity of iysosomal cysteine proteinase during differentiation of rat skeletal muscles. Biochem J ~983;21~871-877. Sohlr I, Fekete E, Yorke F, Cosentino B, Roisen FJ, Bird JWC. Proteinase activities in normal and dystrophic chicken myoblasts in culture. Intracell Prot Catab 198.5;629-63I. Sohar I, Katona G. Regulation of proteinase activities in mammalian tissues. Biol Chem HoppeSeyler 1992;in press.
Further Suggested Reading Hors&h M, KugIer Zs, Dibusz J, Falukiizi J. Nehany megfigyelts Diagnostics Pasteur Miozin IRMA kittel. Laborat Diagnosztika XVII/3, 1990;188(in Hungarian). Tischler ME, Fagan JM, Allen DK. Reduction-oxidation state as a mediator of hormonal control of muscle proteolysis. J Mol Cell Cardiol 1984;16(suppl. 1):36. Stein W. Laboratory diagnosis of acute myocardial infarction. Darmstadt: GIT Verlag, 1988. Kooistra T, Miilard PC, Lloyd JB. Role of thiols in degradation of proteins by cathepsins. B&hem J f 982;204:47I-477.
