Transient membrane association of the precursors of cathepsin C during their transfer into lysosomes

Biochem J. 1991 May 1;275 ( Pt 3)(Pt 3):797-800. doi: 10.1042/bj2750797.

Abstract

Transport of the lysosomal enzyme cathepsin C was studied in Morris hepatoma 7777 cells. Subcellular fractions obtained after isopyenic centrifugation in sucrose gradients of labelled cell homogenates were sequentially extracted by hypo-osmotic shock, Na2CO3 and Triton X-100. Polypeptides related to cathepsin C were immunoprecipitated and analysed by SDS/PAGE and fluorography. At early times after synthesis and for up to 60 min, precursor polypeptides of cathepsin C are distributed in endoplasmic reticulum and Golgi fractions, in membrane-associated form, as Triton X-100 is necessary for their extraction. At 2 h and later after synthesis, intermediate and mature forms of the enzyme can be totally extracted by hypo-osmotic shock from gradient fractions corresponding to the lysosomes of Morris hepatoma 7777 cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biological Transport
  • Cathepsin C
  • Cell Fractionation
  • Cell Membrane / metabolism*
  • Centrifugation, Isopycnic
  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / metabolism*
  • Endoplasmic Reticulum / metabolism
  • Golgi Apparatus / metabolism
  • Immunosorbent Techniques
  • Kinetics
  • Liver Neoplasms, Experimental
  • Lysosomes / metabolism*
  • Protein Precursors / metabolism*
  • Tumor Cells, Cultured

Substances

  • Protein Precursors
  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases
  • Cathepsin C