Isolation, purification and characterization of hemerythrin from Methylococcus capsulatus (Bath).
Earlier work from our laboratory has indicated that a hemerythrin-like protein was over-produced together with the particulate methane monooxygenase (pMMO) when Methylococcus capsulatus (Bath) was grown under high copper concentrations. A homologue of hemerythrin had not previously been found in any prokaryote. To confirm its identity as a hemerythrin, we have isolated and purified this protein by ion-exchange, gel-filtration and hydrophobic interaction chromatography, and characterized it by mass spectrometry, UV-visible, CD, EPR and resonance Raman spectroscopy. On the basis of biophysical and multiple sequence alignment analysis, the protein isolated from M. capsulatus (Bath) is in accord with hemerythrins previously reported from higher organisms. Determination of the Fe content in conjunction with molecular-weight estimation and mass analysis indicates that the native hemerythrin in M. capsulatus (Bath) is a monomer with molecular mass 14.8 kDa, in contrast to hemerythrins from other eukaryotic organisms, where they typically exist as a tetramer or higher oligomers.
Kao
Wei-Chun
WC
Department of Chemistry, National Taiwan University, Taipei 106, Taiwan.
Wang
Vincent C-C
VC
Huang
Yi-Che
YC
Yu
Steve S-F
SS
Chang
Ta-Chau
TC
Chan
Sunney I
SI
D006422:Hemerythrin / Q000302:isolation & purification*; D007501:Iron / Q000032:analysis; D020619:Methylococcus capsulatus / Q000737:chemistry*; D008970:Molecular Weight; D010105:Oxygenases; D013057:Spectrum Analysis
D006422:Hemerythrin; D007501:Iron; D010105:Oxygenases; C021555:methane monooxygenase