Primary structure and unique expression of the 22-kilodalton light chain of human neutrophil cytochrome b.
Cytochrome b comprising 91-kDa and 22-kDa subunits is a critical component of the membrane-bound oxidase of phagocytes that generates superoxide. This important microbicidal system is impaired in inherited disorders known as chronic granulomatous disease (CGD). Previously we determined the sequence of the larger subunit from the cDNA of the CGD gene, the X chromosome locus affected in "X-linked" CGD. To complete the primary structure of the cytochrome b and to assess expression of the smaller subunit, we isolated cDNA clones for the 22-kDa polypeptide by immunoscreening and confirmed their authenticity by direct N-terminal protein sequencing. Although the deduced amino acid sequence of the 22-kDa subunit is not overtly similar to other known cytochromes, we observed a 31-amino acid stretch of 39% identity with polypeptide I of mitochondrial cytochrome c oxidase centered on a potential heme-coordinating histidine. Similarities in the hydropathy profiles and spacing of histidines of the 22-kDa protein and myoglobin suggest structural motifs in common with other heme-containing proteins that are not readily revealed by primary amino acid sequences. Although RNA for the larger subunit has been found only in cells of the phagocytic lineage, stable RNA encoding the 22-kDa subunit was observed in all cell types. However, the stable 22-kDa protein was detected only in phagocytic cells that were expressing the larger subunit RNA. This observation suggests that the large subunit may play a role in regulating the assembly of the heterodimeric cytochrome b.
Parkos
C A
CA
Department of Immunology, Research Institute of the Scripps Clinic, La Jolla, CA 92037.
Dinauer
M C
MC
Walker
L E
LE
Allen
R A
RA
Jesaitis
A J
AJ
Orkin
S H
SH
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003573:Cytochrome b Group / Q000097:blood / Q000235:genetics*; D005796:Genes; D006801:Humans; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009504:Neutrophils / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D014158:Transcription, Genetic
D003573:Cytochrome b Group; D046911:Macromolecular Substances