title: Expression, purification, and characterization of a murine CD4 fragment containing the first two N-terminal domains.
abstract: CD4 is a membrane glycoprotein on T lymphocytes that binds to the same peptide:major histocompatibility complex (MHC) class II molecules recognized by the antigen-specific T cell receptor (TcR). Recent evidence supports the importance of coaggregation of CD4 and TcR for effective T cell activation. Here, we report that a transfected Chinese hamster ovary (CHO) cell line expressing a murine CD4 fragment containing the first two N-terminal domains secretes both monomeric molecules and disulfide-linked multimers. Elimination of the predicted N-linked glycosylation site at residue 161 that is next to the fourth cysteine does not affect the formation of interchain disulfide bonds. N-Terminal amino acid sequencing of the purified CD4 fragment demonstrates that the leader signal sequence is properly cleaved off the expressed protein. Circular dichroism studies suggest that both monomeric and disulfide-linked proteins are folded as primarily beta-sheet.
authors.affiliation: Department of Life Science, National Tsing Hua University, Taiwan, Republic of China. lswwc@life.nthu.edu.tw
authors.forename: W C
authors.initials: WC
authors.lastname: Wang
authors.forename: A A
authors.initials: AA
authors.lastname: Yeh
mesh_terms: D000818:Animals; D015704:CD4 Antigens / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D016466:CHO Cells; D002850:Chromatography, Gel; D002942:Circular Dichroism; D003001:Cloning, Molecular; D006224:Cricetinae; D004220:Disulfides / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D006031:Glycosylation; D051379:Mice; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D011233:Precipitin Tests; D011487:Protein Conformation; D017510:Protein Folding; D021382:Protein Sorting Signals / Q000378:metabolism; D017421:Sequence Analysis; D014162:Transfection
chemical_list: D015704:CD4 Antigens; D004220:Disulfides; D010446:Peptide Fragments; D021382:Protein Sorting Signals