Cardiac troponin T degradation in serum is catalysed by human thrombin.
Cardiac troponin T (cTnT) has been shown to be present in fragmented forms in human serum after acute myocardial infarction (AMI). While calpain-1 and caspase-3 have been identified as intracellular proteases able to cleave the N-terminus of cTnT, it is still unclear which proteases are responsible for the extensive and progressive cTnT fragmentation observed in serum of AMI-patients. In this pilot study we have investigated the possibility that human thrombin may be involved in this process. Purified human cTnT was spiked in unprocessed and deproteinated serum in the presence or absence of either purified human thrombin or PPACK thrombin inhibitor. After immunoprecipitation, SDS-PAGE and Western blotting we observed an increase in cTnT fragmentation when purified thrombin was added to deproteinated serum. Consequently, the addition of thrombin inhibitor to unprocessed serum resulted in a decrease of cTnT fragmentation. Our results suggest that multiple enzymes are involved in cTnT degradation, and that thrombin plays an important role.
Department of Clinical Chemistry, Central Diagnostic Laboratory, Maastricht University Medical Centre, Maastricht, The Netherlands.
Alexander S
AS
Streng
Department of Clinical Chemistry, Central Diagnostic Laboratory, Maastricht University Medical Centre, Maastricht, The Netherlands.
Douwe
D
de Boer
Department of Clinical Chemistry, Central Diagnostic Laboratory, Maastricht University Medical Centre, Maastricht, The Netherlands.
William P T M
WP
van Doorn
Department of Clinical Chemistry, Central Diagnostic Laboratory, Maastricht University Medical Centre, Maastricht, The Netherlands.
Jordy M M
JM
Kocken
Department of Clinical Chemistry, Central Diagnostic Laboratory, Maastricht University Medical Centre, Maastricht, The Netherlands.
Otto
O
Bekers
Department of Clinical Chemistry, Central Diagnostic Laboratory, Maastricht University Medical Centre, Maastricht, The Netherlands. Electronic address: will.wodzig@mumc.nl.
Will K W H
WK
Wodzig
D002384:Catalysis; D006801:Humans; D009206:Myocardium / Q000737:chemistry / Q000378:metabolism; D044967:Serum / Q000737:chemistry* / Q000378:metabolism*; D013917:Thrombin / Q000737:chemistry* / Q000378:metabolism*; D019210:Troponin I / Q000097:blood* / Q000737:chemistry*
D019210:Troponin I; D013917:Thrombin
Cardiac troponin T; Fragmentation; Proteolysis; Thrombin; Western blot