The immunological homology between two filamentous cross-linker phosphoproteins, connectin and cross-bridge region of neurofilament-H, is not affected by the phosphorylation state

J Biochem. 1989 Feb;105(2):226-30. doi: 10.1093/oxfordjournals.jbchem.a122643.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / analysis
  • Antibody Specificity
  • Blotting, Western
  • Cattle
  • Chymotrypsin
  • Connectin
  • Electrophoresis, Polyacrylamide Gel
  • Hydrolysis
  • Intermediate Filament Proteins / analysis
  • Intermediate Filament Proteins / immunology*
  • Male
  • Membranes, Artificial
  • Muscle Proteins / analysis
  • Muscle Proteins / immunology*
  • Muscles / analysis
  • Neurofilament Proteins*
  • Phosphorylation
  • Protein Kinases*
  • Rats
  • Trypsin

Substances

  • Antibodies, Monoclonal
  • Connectin
  • Intermediate Filament Proteins
  • Membranes, Artificial
  • Muscle Proteins
  • Neurofilament Proteins
  • neurofilament protein H
  • Protein Kinases
  • Chymotrypsin
  • Trypsin