Activity-based protein profiling of intraoperative serine hydrolase activities during cardiac surgery
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Date
2018-09-07
Authors
Navarette, Mario
Ho, Julie
Dwivedi, Ravi
Choi, Nora
Ezzati, Peyman
Krokhin, Oleg
Spicer, Vic
Arora, Rakesh
Rigatto, Claudio
Wilkins, John
Journal Title
Journal ISSN
Volume Title
Publisher
American Chemical Society Publications
Abstract
The processes involved in the initiation of acute kidney injury (AKI) following cardiopulmonary bypass (CPB) are thought to occur during the intraoperative period. Such a rapid development might indicate that some of the inductive events are not
dependent on de novo protein synthesis, raising the possibility that changes in activities of
pre-existing enzymes could contribute to the development of AKI. Activity-based protein
profiling (ABPP) was used to compare the serine hydrolase enzyme activities present in the urines of CPB patients who subsequently developed AKI versus those who did not (non-AKI) during the intra- and immediate postoperative periods. Sequential urines collected from a nested case-control cohort of AKI and non-AKI patients were reacted with a serine hydrolase activity probe, fluorophosphonate-TAMRA, and separated by
SDS-PAGE. The patterns and levels of probe-labeled proteins in the two groups were initially comparable. However, within 1 h of CPB there were significant pattern changes in the AKI group. Affinity purification and mass spectrometry-based analysis of
probe-labeled enzymes in AKI urines at 1 h CPB and arrival to the intensive care unit (ICU) identified 28 enzymes. Quantitative analysis of the activity of one of the identified enzymes, kallikrein-1, revealed some trends suggesting differences in the levels and temporal patterns of enzyme activity between a subset of patients who developed AKI and those who did not. A comparative analysis of affinity-purified probe reacted urinary proteins from these patient groups during the intraoperative period suggested the presence of both shared and unique enzyme patterns. These results indicate that there are intraoperative changes in the levels and types of serine hydrolase activities in patients who subsequently develop AKI. However, the role of these activity differences in the development of AKI remains to be determined.
Description
Keywords
activity-based protein profiling, acute kidney injury, proteomics, serine hydrolase, kallikrein-1, cardiopulmonary bypass
Citation
J Proteome Res 17(10): 3547-3556, 2018