Protein surface charge effect on 3D domain swapping in cells for c-type cytochromes.
Many c-type cytochromes (cyts) can form domain-swapped oligomers. The positively charged Hydrogenobacter thermophilus (HT) cytochrome (cyt) c552 forms domain-swapped oligomers during expression in the Escherichia coli (E. coli) expression system, but the factors influencing the oligomerization remain unrevealed. Here, we found that the dimer of the negatively charged Shewanella violacea (SV) cyt c5 exhibits a domain-swapped structure, in which the N-terminal helix is exchanged between protomers, similar to the structures of the HT cyt c552 and Pseudomonas aeruginosa (PA) cyt c551 domain-swapped dimers. Positively charged horse cyt c and HT cyt c552 domain swapped during expression in E. coli, whereas negatively charged PA cyt c551 and SV cyt c5 did not. Oligomers were formed during expression in E. coli for HT cyt c552 attached to either a co- or post-translational signal peptide for transportation through the cytoplasm membrane, but not for PA cyt c551 attached to either signal peptide. HT cyt c552 formed oligomers in E. coli in the presence and absence of rare codons. More oligomers were obtained from the in vitro folding of horse cyt c and HT cyt c552 by the addition of negatively charged liposomes during folding, whereas the amount of oligomers for the in vitro folding of PA cyt c551 and SV cyt c5 did not change significantly by the addition. These results indicate that the protein surface charge affects the oligomerization of c-type cyts in cells; positively charged c-type cyts assemble on a negatively charged membrane, inducing formation of domain-swapped oligomers during folding.
['Yang|Hongxu|H|', 'Yamanaka|Masaru|M|', 'Nagao|Satoshi|S|', 'Yasuhara|Kazuma|K|', 'Shibata|Naoki|N|', 'Higuchi|Yoshiki|Y|', 'Hirota|Shun|S|']
D001426:Bacterial Proteins D003574:Cytochrome c Group D004926:Escherichia coli D000072417:Protein Domains D055503:Protein Multimerization D011550:Pseudomonas aeruginosa D020592:Shewanella D013499:Surface Properties
Protein surface charge Domain swapping c-type cytochrome Oligomerization Lipid-protein interaction