Thiol-based regulation of glyceraldehyde-3-phosphate dehydrogenase in blood bank-stored red blood cells: a strategy to counteract oxidative stress.
Author(s): Rinalducci S, Marrocco C, Zolla L
Publication: Transfusion, 2015, Vol. 55, Page 499-506
PubMed ID: 25196942 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to determine the effects of refrigerated storage on glyceraldehyde-3-phosphate dehydrogenase (GAPDH) levels and molecular modification in red blood cell (RBC) membranes and cytosol.
Conclusion of Paper
During storage of RBCs, GAPDH moved from the RBC membrane to the cytosol and then back to the membrane. In addition, on day 21 of storage, cytosolic GAPDH had an intramolecular disulfide bond that was not present on day 0 which, the authors postulate, may cause a reversible inactivation of the molecule.
Studies
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Study Purpose
The purpose of this study was to determine the effects of refrigerated storage on GAPDH levels and molecular modification in RBC membranes and cytosol. Leukoreduced RBCs in saline-adenine-glucose-mannitol (SAGM) were obtained from 5 healthy donors and stored at 4°C.
Summary of Findings:
Cytosolic GAPDH activity increased rapidly between day 0 and day 7 of refrigerated storage of RBCs (p<0.001) and was slightly higher on day 14 than day 7 of storage but was lower on day 21 than on day 0 (p<0.001). A slight increase in GAPDH activity was observed between day 21 and day 42 of RBC storage such that on day 42, GAPDH activity was slightly higher than on day 0 (p<0.01). Western blot analysis showed that membrane bound GAPDH decreased while cytosolic GAPDH increased during the first 14 days of refrigerated storage of RBCs (p<0.001, both), but that after 28 days, levels of GAPDH in the cytosol were comparable to those on day 0, and levels of GAPDH in the membranes were higher than on day 0 (p<0.001). Under non-reducing conditions, no GAPDH aggregates were observed in the cytosolic or membrane fractions, but an intramolecular disulfide bond between cysteine 152 and 156 was found in cytosolic GAPDH on day 21, but not day 0 of storage. The authors state this intramolecular disulfide bond may cause a reversible inactivation of GAPDH.
Biospecimens
Preservative Types
- Other Preservative
Diagnoses:
- Normal
Platform:
Analyte Technology Platform Protein LC-ESI-MS/MS Protein Colorimetric assay Protein Western blot Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Western blot Specific Targeted peptide/protein GAPDH
Storage Storage duration 0 days
7 days
14 days
21 days
28 days
35 days
42 days