About 14 proteins were tested for specific oxidative scission catalyzed by metal ions in the presence of ascorbate and oxidizing agents (O2 or hydrogen peroxide). Only four of them were degraded by Fe3+/Fe2+- ascorbate, twelve – by Cu2+/Cu+-ascorbate and two proteins (α- and β-caseins) were degraded by Pd2+ ions. The rate and the intensity of degradation are very different for various proteins. For the most of tested proteins only a small fraction of molecules was degraded. None of them was degraded completely. Two possible reasons of protein stability against oxidative degradation may be proposed as follows: either there is no metal binding site in a protein molecule, or metal binding ligands of protein undergo a rapid oxidative modification and the metal ion is released from the binding site. Human growth hormone was cut specifically at two sites by Cu2+/Cu+-ascorbate system. At least one of amino acid residues of this protein was modified by formation of reactive carbonyl.
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Abbreviations
- BSA:
-
bovine serum albumin
- CAPS:
-
3-(cyclohexylamino) propanesulfonic acid
- Cu/asc:
-
Cu2+/Cu+-ascorbate
- DNPH:
-
2,4-dinitrophenylhydrazine
- DTT:
-
dithiothreitol
- Fe/asc:
-
Fe3+/Fe2+-ascorbate
- GCSF:
-
recombinant human granulocyte colony-stimulating factor
- GuHCl:
-
guanidinium hydrochloride
- hGH:
-
recombinant human growth hormone
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- TCA:
-
trichloroacetic acid
- TFA:
-
trifluoroacetic acid
- TNF:
-
recombinant human tumor necrosis factor-alpha
- Tricine:
-
N-[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]glycine
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Sereikaite, J., Jachno, J., Santockyte, R. et al. Protein Scission by Metal Ion–Ascorbate System. Protein J 25, 369–378 (2006). https://doi.org/10.1007/s10930-006-9014-7
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DOI: https://doi.org/10.1007/s10930-006-9014-7