Research Article Open Access

Protective Effects of Covalent Cross-Linking on Proteolysis of Human Coproporphyrinogen Oxidase and Implications for Porphyria

Ahmed W. Jafri1, Jason R. Stephenson1, Justin B. Morgenthaler1, Jon A. Friesen1 and Marjorie A. Jones1
  • 1 Illinois State University, United States

Abstract

The effects of covalent cross-linkers on the enzyme, coproporphyrinogen oxidase, had been previously studied but their role in protecting the enzyme from protease cleavage has not been evaluated. Therefore, we examined how the cross-linker bis (sulfosuccinimidyl) suberate (BS3) affects the ability of trypsin to digest purified, wild type recombinant human coproporphyrinogen oxidase and selected mutants. Following incubation, the apparent molecular weights of peptides were evaluated by SDS-PAGE and enzymatic activity was assessed by spectroscopy following HPLC. For both wild type and mutants, the results indicated that the cross-linker was indeed able to protect against trypsin digestion relative to the enzyme incubated with trypsin in the absence of the cross-linker. These data have implications for the episodic nature of porphyria.

American Journal of Biochemistry and Biotechnology
Volume 4 No. 4, 2008, 442-449

DOI: https://doi.org/10.3844/ajbbsp.2008.442.449

Submitted On: 15 July 2008 Published On: 31 December 2008

How to Cite: Jafri, A. W., Stephenson, J. R., Morgenthaler, J. B., Friesen, J. A. & Jones, M. A. (2008). Protective Effects of Covalent Cross-Linking on Proteolysis of Human Coproporphyrinogen Oxidase and Implications for Porphyria. American Journal of Biochemistry and Biotechnology, 4(4), 442-449. https://doi.org/10.3844/ajbbsp.2008.442.449

  • 2,929 Views
  • 2,357 Downloads
  • 0 Citations

Download

Keywords

  • Coproporphyrinogen oxidase
  • BS3
  • cross-linking
  • trypsin
  • porphyria