Open AccessUse of Cross-linking to Assess Subunit Interaction of Recombinant Human Coproporphyrinogen Oxidase
Author(s) -
Jason R. Stephenson,
Nancy E. Thomas,
Jon A. Friesen,
Marjorie A. Jones
Publication year - 2005
Publication title -
american journal of biochemistry and biotechnology/american journal of biochemistry and biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.161
H-Index - 31
eISSN - 1553-3468
pISSN - 1558-6332
DOI - 10.3844/ajbbsp.2005.103.106
Subject(s) - protein subunit , recombinant dna , chemistry , computational biology , biochemistry , biology , gene
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