Research Article Open Access

Use of Cross-linking to Assess Subunit Interaction of Recombinant Human Coproporphyrinogen Oxidase

Jason R. Stephenson1, Nancy E. Thomas1, Jon A. Friesen1 and Marjorie A. Jones1
  • 1 Illinois State University, United States

Abstract

To provide further evidence for a dimeric form of coproporphyrinogen oxidase reported using the conventional hydrodynamic methods, bifunctional cross-linkers were incubated with purified, recombinant human coproporphyrinogen oxidase to determine subunit interaction in solution. The use of cross-linkers provides an effective way to demonstrate subunit association and allows for assessment of activity upon covalent cross-linking. Following incubation with selected cross-linkers, enzyme apparent molecular weight was evaluated using SDS-PAGE and enzymatic activity was monitored by spectroscopy following HPLC. The predominate multimeric form of coproporphyrinogen oxidase observed had a mass that corresponded to a dimer, indicating that coproporphyrinogen oxidase most likely functions as a homodimer in solution.

American Journal of Biochemistry and Biotechnology
Volume 1 No. 2, 2005, 103-106

DOI: https://doi.org/10.3844/ajbbsp.2005.103.106

Submitted On: 28 July 2005 Published On: 30 June 2005

How to Cite: Stephenson, J. R., Thomas, N. E., Friesen, J. A. & Jones, M. A. (2005). Use of Cross-linking to Assess Subunit Interaction of Recombinant Human Coproporphyrinogen Oxidase. American Journal of Biochemistry and Biotechnology, 1(2), 103-106. https://doi.org/10.3844/ajbbsp.2005.103.106

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Keywords

  • Coproporphyrinogen oxidase
  • cross-linking
  • multimeric
  • protein structure