Monoclonal antibodies specific to the integral membrane protein P0 of bovine peripheral nerve myelin.
Two monoclonal antibodies (mAbs), 58A and 46E, were generated against the major protein P0 of bovine peripheral nervous system myelin (PNSM). The reactivities of the mAbs were assessed by enzyme-linked immunosorbent assay (ELISA), Western blot, and immunohistochemistry. Both mAbs, 58A and 46E, reacted to PNSM of bovine, human, rat and rabbit, but not to chicken PNSM or the brains of rat and rabbit. In the Western blot, these mAbs showed specific binding to bovine P0 as well as deglycosylated P0, but not to myelin-associated glycoprotein (MAG) of bovine spinal cord. The analyses of the lysylendopeptidase-digested peptides of bovine P0 revealed that the epitopes for the mAbs 58A and 46E were located on the amino acid residues 68-79 and 210-216, respectively. Since the mAbs 58A and 46E recognize the extracellular domain and the cytoplasmic domain of P0, respectively, they could be useful for studies on P0's role in myelin formation, its adhesive properties, and functions of the N-terminal extracellular and C-terminal cytoplasmic domains of the protein.
Yoshimura
K
K
Dept. of Physiology, Saitama Medical School, Japan.
Negishi
T
T
Kaneko
A
A
Sakamoto
Y
Y
Kitamura
K
K
Hosokawa
T
T
Hamaguchi
K
K
Nomura
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002417:Cattle; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D006801:Humans; D006825:Hybridomas; D007132:Immunoglobulin Isotypes / Q000032:analysis; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009101:Multiple Myeloma; D018993:Myelin P0 Protein / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D009186:Myelin Sheath / Q000737:chemistry*; D010525:Peripheral Nerves / Q000737:chemistry* / Q000648:ultrastructure; D017434:Protein Structure, Tertiary; D011817:Rabbits; D051381:Rats; D014407:Tumor Cells, Cultured / Q000737:chemistry
D000911:Antibodies, Monoclonal; D007132:Immunoglobulin Isotypes; D018993:Myelin P0 Protein