Cloning and expression of mouse legumain, a lysosomal endopeptidase

Biochem J. 1998 Oct 1;335 ( Pt 1)(Pt 1):111-7. doi: 10.1042/bj3350111.

Abstract

Legumain, a recently discovered mammalian cysteine endopeptidase, was found in all mouse tissues examined, but was particularly abundant in kidney and placenta. The distribution in subcellular fractions of mouse and rat kidney showed a lysosomal localization, and activity was detectable only after the organelles were disrupted. Nevertheless, ratios of legumain activity to that of cathepsin B differed considerably between mouse tissues. cDNA encoding mouse legumain was cloned and sequenced, the deduced amino acid sequence proving to be 83% identical to that of the human protein [Chen, Dando, Rawlings, Brown, Young, Stevens, Hewitt, Watts and Barrett (1997) J. Biol. Chem. 272, 8090-8098]. Recombinant mouse legumain was expressed in human embryonic kidney 293 cells by use of a vector containing a cytomegalovirus promoter. The recombinant enzyme was partially purified and found to be an asparagine-specific endopeptidase closely similar to naturally occurring pig kidney legumain.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cathepsin B / metabolism
  • Cells, Cultured
  • Cloning, Molecular
  • Cysteine Endopeptidases / biosynthesis*
  • Cysteine Endopeptidases / genetics*
  • DNA, Complementary / metabolism
  • Databases, Factual
  • Humans
  • Kidney / chemistry
  • Mice
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • Plant Proteins*
  • Rats
  • Recombinant Proteins / metabolism
  • Sequence Analysis, DNA
  • Swine

Substances

  • DNA, Complementary
  • Plant Proteins
  • Recombinant Proteins
  • Cysteine Endopeptidases
  • Cathepsin B
  • asparaginylendopeptidase

Associated data

  • GENBANK/AJ000990