The immunological homology between two filamentous cross-linker phosphoproteins, connectin and cross-bridge region of neurofilament-H, is not affected by the phosphorylation state.
It has recently been shown that a monoclonal antibody SM 1-36-2 against connectin, an elastic filament of striated muscles, binds to the "elastic" domain of the molecule, and that the H subunit of neurofilament (NF-H), an intermediate filament of nerve cells, shares a homologous domain (Shimizu, T. et al. (1988) Biomed. Res. 9, 227-234 and Itoh, Y. et al. (1988) J. Biochem. 104, 504-508). In order to characterize (1) the intramolecular localization of the domain in the NF-H and (2) the effect of the phosphorylation state on the immunoreactivity, the homologous domain in the NF-H was analyzed by Western blotting after limited digestion with trypsin or alpha-chymotrypsin and dephosphorylation with E. coli alkaline phosphatase. It was found that (1) the epitope was located not in the core region but in the carboxyl-terminal peripheral (cross-bridge) region of NF-H and (2) the epitopes in connectin and NF-H were not affected by the phosphorylation state.
Matsumura
K
K
Department of Neurology, Faculty of Medicine, University of Tokyo.
Shimizu
T
T
Mannen
T
T
Maruyama
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D000918:Antibody Specificity; D015153:Blotting, Western; D002417:Cattle; D002918:Chymotrypsin; D064211:Connectin; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D007381:Intermediate Filament Proteins / Q000032:analysis / Q000276:immunology*; D008297:Male; D008567:Membranes, Artificial; D009124:Muscle Proteins / Q000032:analysis / Q000276:immunology*; D009132:Muscles / Q000032:analysis; D016900:Neurofilament Proteins; D010766:Phosphorylation; D011494:Protein Kinases; D051381:Rats; D014357:Trypsin
D000911:Antibodies, Monoclonal; D064211:Connectin; D007381:Intermediate Filament Proteins; D008567:Membranes, Artificial; D009124:Muscle Proteins; D016900:Neurofilament Proteins; C051774:neurofilament protein H; D011494:Protein Kinases; D002918:Chymotrypsin; D014357:Trypsin