Protein-Chemical Identification of the Major Cleavage Sites of the Ca2⊕Proteinase on Murine Vimentin, the Mesenchymal Intermediate Filament Protein
- 1 January 1986
- journal article
- research article
- Published by Walter de Gruyter GmbH in Biological Chemistry Hoppe-Seyler
- Vol. 367 (2), 1147-1152
- https://doi.org/10.1515/bchm3.1986.367.2.1147
Abstract
Neutral thiol proteinases (calpains), activated by calcium are involved in the intracellular turnover of intermediate filaments but the precise position of the cleavage points has remained unknown. Here we identify by direct sequence analysis the major cleavage sites found when murine vimentin is digested by limited proteolysis in vitro with calpain purified from porcine kidney. Contrary to some previous suggestions, no absolute sequence specificity could be detected although 10 specific sites have been identified. This result is in line with cDNA derived amino-acid sequence of a calpain, which pointed to a similarity of the catalytic site with the active sites in papain, cathepsin and actinidin. However, all major cleavage sites are located within regions of the vimentin molecule, which in current models of intermediate filament structure are thought to be non-helical: the amino-terminal headpiece, the carboxyl-terminal tail-piece and the spacer separating the two major coiled-coil domains. The sequence information about the cleavage sites was extended to provide the amino-terminal 119 residues of murine vimentin.This publication has 22 references indexed in Scilit:
- An Immunoblot Study of Neurofilament Degradation In Situ and During Calcium‐Activated ProteolysisJournal of Neurochemistry, 1985
- Large scale isolation, purification, and partial characterization of the intermediate filament-specific, Ca2+-activated proteinase from porcine kidney and Ehrlich ascites tumor cells: A comparative studyArchives of Biochemistry and Biophysics, 1984
- The amino acid sequence of chicken muscle desmin provides a common structural model for intermediate filament proteins.The EMBO Journal, 1982
- Proteinchemical characterization of three structurally distinct domains along the protofilament unit of desmin 10 nm filamentsCell, 1982
- A rapid method for the large scale purification of the intermediate filament protein vimentin by single-stranded DNA-cellulose affinity chromatographyBiochemical and Biophysical Research Communications, 1982
- A gas-liquid solid phase peptide and protein sequenator.Journal of Biological Chemistry, 1981
- Properties of a Ca2+‐Activated Protease Specific for the Intermediate‐Sized Filament Protein Vimentin in Ehrlich‐Ascites‐Tumour CellsEuropean Journal of Biochemistry, 1981
- Studies of a Calcium-Activated Neutral Protease from Chicken Skeletal MuscleThe Journal of Biochemistry, 1979
- A novel manual method for protein-sequence analysisBiochemical Journal, 1976