Use of Cross-linking to Assess Subunit Interaction of Recombinant Human Coproporphyrinogen Oxidase
Jason R. Stephenson, Nancy E. Thomas, Jon A. Friesen and Marjorie A. Jones
DOI : 10.3844/ajbbsp.2005.103.106
American Journal of Biochemistry and Biotechnology
Volume 1, Issue 2
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.
© 2005 Jason R. Stephenson, Nancy E. Thomas, Jon A. Friesen and Marjorie A. Jones. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.