Expression of a mouse-channel catfish chimeric IgM molecule in a mouse myeloma cell.
Fusion genes encoding a murine VH domain and the constant region domains of the mu chain from the channel catfish, Ictalurus punctatus, were stably expressed in the lambda light chain producing mouse myeloma cell line J558L. Although the pathways of pre-mRNA processing for expression of membrane (micron and secreted (microsecond) forms of the mu chain differ between mammals and teleosts, mRNAs encoding both catfish micron and microsecond were correctly expressed in the mouse myeloma cells. The mouse-channel catfish chimeric mu chain polypeptide was able to associate covalently with the mouse lambda light chain and assemble, intracellularly, into polymers of covalent structure (microL)2-8 which resembled those seen with native catfish IgM. In contrast to native catfish IgM, the mouse-catfish chimeric IgM showed the property of binding strongly to protein A of Staphylococcus aureus. The mouse-channel catfish chimeric IgM was core-glycosylated, but did not contain terminal sialic acid. Secretion rates for the chimeric IgM were low, and the possibility could not be excluded that extracellular chimeric IgM was released from dead or dying cells. The reason(s) for the intracellular retention of the chimeric IgM (probably in the endoplasmic reticulum) are not known, but those mechanisms involving retention via cysteine residues were excluded.
Ledford
B E
BE
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston 29425-2211.
Magor
B G
BG
Middleton
D L
DL
Miller
R L
RL
Wilson
M R
MR
Miller
N W
NW
Clem
L W
LW
Warr
G W
GW
D000818:Animals; D001483:Base Sequence; D002678:Chimera; D015870:Gene Expression; D007059:Ictaluridae; D007075:Immunoglobulin M / Q000235:genetics / Q000378:metabolism*; D007146:Immunoglobulin lambda-Chains; D007148:Immunoglobulin mu-Chains; D051379:Mice; D008969:Molecular Sequence Data; D010954:Plasmacytoma / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis; D016415:Sequence Alignment; D013205:Staphylococcal Protein A / Q000378:metabolism; D013462:Sulfur Radioisotopes; D014162:Transfection; D014407:Tumor Cells, Cultured
D007075:Immunoglobulin M; D007146:Immunoglobulin lambda-Chains; D007148:Immunoglobulin mu-Chains; D012333:RNA, Messenger; D013205:Staphylococcal Protein A; D013462:Sulfur Radioisotopes