A major theme of TBI (traumatic brain injury) pathology is the over-activation of multiple proteases. We have previously shown that calpain-1 and -2, and caspase-3 simultaneously produced αII-spectrin BDPs (breakdown products) following TBI. In the present study, we attempted to identify a comprehensive set of protease substrates (degradome) for calpains and caspase-3. We further hypothesized that the TBI differential proteome is likely to overlap significantly with the calpain- and caspase-3-degradomes. Using a novel HTPI (high throughput immunoblotting) approach and 1000 monoclonal antibodies (PowerBlot™), we compared rat hippocampal lysates from 4 treatment groups: (i) naïve, (ii) TBI (48 h after controlled cortical impact), (iii) in vitro calpain-2 digestion and (iv) in vitro caspase-3 digestion. In total, we identified 54 and 38 proteins that were vulnerable to calpain-2 and caspase-3 proteolysis respectively. In addition, the expression of 48 proteins was down-regulated following TBI, whereas that of only 9 was up-regulated. Among the proteins down-regulated in TBI, 42 of them overlapped with the calpain-2 and/or caspase-3 degradomes, suggesting that they might be proteolytic targets after TBI. We further confirmed several novel TBI-linked proteolytic substrates, including βII-spectrin, striatin, synaptotagmin-1, synaptojanin-1 and NSF (N-ethylmaleimide-sensitive fusion protein) by traditional immunoblotting. In summary, we demonstrated that HTPI is a novel and powerful method for studying proteolytic pathways in vivo and in vitro.
Skip Nav Destination
Article navigation
March 2006
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
- PDF Icon PDF LinkEditorial Board
Research Article|
February 24 2006
Comparing calpain- and caspase-3-mediated degradation patterns in traumatic brain injury by differential proteome analysis
Ming Cheng Liu;
Ming Cheng Liu
*Center for Neuroproteomics and Biomarkers Research, Department of Psychiatry, McKnight Brain Institute, University of Florida, P.O. Box 100256, Gainesville, FL 32610, U.S.A.
†Center for Traumatic Brain Injury Studies, Department of Neuroscience, McKnight Brain Institute, University of Florida, P.O. Box 100256, Gainesville, FL 32610, U.S.A.
Search for other works by this author on:
Veronica Akle;
Veronica Akle
*Center for Neuroproteomics and Biomarkers Research, Department of Psychiatry, McKnight Brain Institute, University of Florida, P.O. Box 100256, Gainesville, FL 32610, U.S.A.
†Center for Traumatic Brain Injury Studies, Department of Neuroscience, McKnight Brain Institute, University of Florida, P.O. Box 100256, Gainesville, FL 32610, U.S.A.
Search for other works by this author on:
Wenrong Zheng;
Wenrong Zheng
*Center for Neuroproteomics and Biomarkers Research, Department of Psychiatry, McKnight Brain Institute, University of Florida, P.O. Box 100256, Gainesville, FL 32610, U.S.A.
†Center for Traumatic Brain Injury Studies, Department of Neuroscience, McKnight Brain Institute, University of Florida, P.O. Box 100256, Gainesville, FL 32610, U.S.A.
Search for other works by this author on:
Jitendra R. Dave;
Jitendra R. Dave
‡Department of Neuropharmacology and Molecular Biology, Division of Neurosciences, Walter Reed Army Institute of Research, Silver Spring, MD, U.S.A.
Search for other works by this author on:
Frank C. Tortella;
Frank C. Tortella
‡Department of Neuropharmacology and Molecular Biology, Division of Neurosciences, Walter Reed Army Institute of Research, Silver Spring, MD, U.S.A.
Search for other works by this author on:
Ronald L. Hayes;
Ronald L. Hayes
*Center for Neuroproteomics and Biomarkers Research, Department of Psychiatry, McKnight Brain Institute, University of Florida, P.O. Box 100256, Gainesville, FL 32610, U.S.A.
†Center for Traumatic Brain Injury Studies, Department of Neuroscience, McKnight Brain Institute, University of Florida, P.O. Box 100256, Gainesville, FL 32610, U.S.A.
§Banyan Biomarkers, Inc. 12085 Research Drive, Suite 180, Alachua, FL 32615, U.S.A.
Search for other works by this author on:
Kevin K. W. Wang
Kevin K. W. Wang
1
*Center for Neuroproteomics and Biomarkers Research, Department of Psychiatry, McKnight Brain Institute, University of Florida, P.O. Box 100256, Gainesville, FL 32610, U.S.A.
†Center for Traumatic Brain Injury Studies, Department of Neuroscience, McKnight Brain Institute, University of Florida, P.O. Box 100256, Gainesville, FL 32610, U.S.A.
§Banyan Biomarkers, Inc. 12085 Research Drive, Suite 180, Alachua, FL 32615, U.S.A.
1To whom correspondence should be addressed (email kwang1@ufl.edu).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
June 06 2005
Revision Received:
December 13 2005
Accepted:
December 14 2005
Accepted Manuscript online:
December 14 2005
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2006
Biochem J (2006) 394 (3): 715–725.
Article history
Received:
June 06 2005
Revision Received:
December 13 2005
Accepted:
December 14 2005
Accepted Manuscript online:
December 14 2005
Citation
Ming Cheng Liu, Veronica Akle, Wenrong Zheng, Jitendra R. Dave, Frank C. Tortella, Ronald L. Hayes, Kevin K. W. Wang; Comparing calpain- and caspase-3-mediated degradation patterns in traumatic brain injury by differential proteome analysis. Biochem J 15 March 2006; 394 (3): 715–725. doi: https://doi.org/10.1042/BJ20050905
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.