Structure of recombinant human interleukin 5 produced by Chinese hamster ovary cells.
The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. Each peptide fragment generated by Achromobacter protease I (API) digestion was purified and characterized by amino acid analysis and amino acid sequence analysis. After digestion with API, we could identify all the peptides which were expected from human IL-5 cDNA sequence. The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. In addition, we concluded that Thr-3 and Asn-28 were glycosylated. The results indicate that primary structure of rhIL-5 is highly homogeneous and observed heterogeneity is due to the difference in the content of carbohydrate.
Minamitake
Y
Y
Suntory Institute for Biomedical Research, Osaka.
Kodama
S
S
Katayama
T
T
Adachi
H
H
Tanaka
S
S
Tsujimoto
M
M
D000595:Amino Acid Sequence; D000818:Animals; D006224:Cricetinae; D004220:Disulfides; D005260:Female; D006031:Glycosylation; D015848:Interleukin-5 / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010053:Ovary / Q000166:cytology; D010449:Peptide Mapping; D011994:Recombinant Proteins / Q000302:isolation & purification*
D004220:Disulfides; D015848:Interleukin-5; D011994:Recombinant Proteins