Characterization of recombinant human interleukin 4 receptor from CHO cells: role of N-linked oligosaccharides.
Interleukin 4 (IL-4) mediates its biological activities through interaction with its receptor on the cell surface. A recombinant extracellular domain of the alpha subunit of human interleukin 4 receptor was expressed in CHO cells and purified to homogeneity by a combination of ion exchange and immunoaffinity chromatography. Analysis of the purified protein by MALDI MS provided an average mass of 38,241 Da while microsequencing identified the site of the signal sequence processing to be Ser23-Gly24. The receptor was highly glycosylated, containing N-linked complex oligosaccharides with bi-, tri-, and tetraantennary structures. Five of the six potential glycosylation sites could be assigned to Asn residues 53, 98, 128, 134 and 176. N-deglycosylation increased aggregation and reduced solubility of the receptor but did not affect its IL-4 binding activity. These observations provide preliminary insights into the role of N-linked oligosaccharides in IL-4 receptor biosynthesis and function at the cell surface.
Rajan
N
N
Schering-Plough Research Institute, Kenilworth, New Jersey 07033.
Tsarbopoulos
A
A
Kumarasamy
R
R
O'Donnell
R
R
Taremi
S S
SS
Baldwin
S W
SW
Seelig
G F
GF
Fan
X
X
Pramanik
B
B
Le
H V
HV
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002241:Carbohydrates / Q000032:analysis; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D006224:Cricetinae; D006020:Glycopeptides / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D015847:Interleukin-4 / Q000378:metabolism; D046911:Macromolecular Substances; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification; D018123:Receptors, Interleukin / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D019948:Receptors, Interleukin-4; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification
D002241:Carbohydrates; D006020:Glycopeptides; D046911:Macromolecular Substances; D009844:Oligosaccharides; D018123:Receptors, Interleukin; D019948:Receptors, Interleukin-4; D011994:Recombinant Proteins; D015847:Interleukin-4