Synthetic peptides corresponding to various hydrophilic regions of the large subunit of cytochrome b558 inhibit superoxide generation in a cell-free system from neutrophils.
Cytochrome b558 is a component of the superoxide-generating system in neutrophils and plays key roles in both the assembly of a functional complex with cytosolic proteins and shuttling an electron from NADPH to molecular oxygen. To determine the role of predicted hydrophilic domains of gp91-phox, a glycosylated subunit of the cytochrome, we synthesized peptides corresponding to the regions and tested whether they affected superoxide generation in the cell-free system obtained from human neutrophils. Among twelve peptides tested, six peptides, four of which correspond to previously unreported regions, inhibited superoxide generation in the cell-free system. All of the active peptides were effective when added to the system before activation with sodium dodecyl sulfate. Four peptides, including two peptides corresponding to two newly identified regions, inhibited the translocation of the cytosolic components, p47-phox and p67-phox. The extent of inhibition on translocation of these components varied depending on the peptide used.
Park
M Y
MY
The Institute of Medical Science, University of Tokyo, Minatoku, Japan.
Imajoh-Ohmi
S
S
Nunoi
H
H
Kanegasaki
S
S
D000595:Amino Acid Sequence; D001693:Biological Transport, Active / Q000187:drug effects; D002474:Cell-Free System; D003573:Cytochrome b Group / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D003600:Cytosol / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D008969:Molecular Sequence Data; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism*; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000235:genetics / Q000494:pharmacology*; D010750:Phosphoproteins / Q000378:metabolism; D011487:Protein Conformation; D013481:Superoxides / Q000378:metabolism*
D003573:Cytochrome b Group; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D010750:Phosphoproteins; C079789:neutrophil cytosol factor 67K; D013481:Superoxides; C036888:cytochrome b558; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1