Proteomic and phosphoproteomic analyses of the soluble fraction following acute spinal cord contusion in rats.
Traumatic spinal cord injury (SCI) causes marked neuropathological changes in the spinal cord, resulting in limited functional recovery. Currently, there are no effective treatments, and the mechanisms underlying these neuropathological changes are not completely understood. In this study, two-dimensional gel electrophoresis coupled with mass spectrometry was used to investigate injury-related changes in the abundance (SYPRO Ruby stain) and phosphorylation (Pro-Q Diamond stain) of proteins from the soluble fraction of the lesion epicenter at 24 h following SCI. Over 1500 SYPRO Ruby-stained spots and 100 Pro-Q Diamond-stained spots were examined. We identified 26 unique proteins within 38 gel spots that differentially changed in abundance, phosphorylation, or both in response to SCI. Protein redundancies among the gel spots were likely due to differences in proteolysis, post-translational modifications, and the existence of isoforms. The proteins affected were blood-related proteins, heat-shock proteins, glycolytic enzymes, antioxidants, and proteins that function in cell structure, cell signaling, DNA damage, and protein degradation. These protein changes post injury may suggest additional avenues of investigation into the underlying molecular mechanisms responsible for the pathophysiological consequences of SCI.
Chen
Anshu
A
University of Kentucky, Department of Physical Medicine and Rehabilitation, Lexington, Kentucky 40536-0509, USA.
McEwen
Melanie L
ML
Sun
Shixin
S
Ravikumar
Rangaswamyrao
R
Springer
Joe E
JE
D000818:Animals; D004129:Dimethylpolysiloxanes; D004195:Disease Models, Animal; D015180:Electrophoresis, Gel, Two-Dimensional; D004734:Energy Metabolism / Q000502:physiology; D005260:Female; D005990:Glycerol; D006470:Hemorrhage / Q000378:metabolism / Q000503:physiopathology; D013058:Mass Spectrometry; D009410:Nerve Degeneration / Q000378:metabolism / Q000503:physiopathology; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000378:metabolism*; D009942:Organometallic Compounds; D010750:Phosphoproteins / Q000032:analysis / Q000378:metabolism; D010766:Phosphorylation; D040901:Proteomics / Q000379:methods*; D051381:Rats; D020318:Rats, Long-Evans; D015398:Signal Transduction / Q000502:physiology; D013116:Spinal Cord / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D013119:Spinal Cord Injuries / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D013194:Staining and Labeling; D013347:Subcellular Fractions / Q000378:metabolism; D013997:Time Factors
D004129:Dimethylpolysiloxanes; D009419:Nerve Tissue Proteins; D009942:Organometallic Compounds; D010750:Phosphoproteins; C118954:Pro-Q aerosol foam; C515601:Sypro Ruby; D005990:Glycerol
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